Pyrido[2,3-b]pyrazin-8-substituted Compounds and Their Use

ABSTRACT

The present invention pertains generally to the field of therapeutic compounds for treating proliferative disorders, cancer, etc., and more specifically to certain pyrido[2,3-b]pyrazin-8-substituted compounds, as described herein, which, inter alia, inhibit RAF (e.g., B-RAF) activity. The present invention also pertains to pharmaceutical compositions comprising such compounds, and the use of such compounds and compositions, both in vitro and in vivo, to inhibit RAF (e.g., BRAF) activity, to inhibit receptor tyrosine kinase (RTK) activity, to inhibit cell proliferation, and in the treatment of diseases and disorders that are ameliorated by the inhibition of RAF, RTK, etc., proliferative disorders such as cancer (e.g., colorectal cancer, melanoma), etc.

RELATED APPLICATION

This application is a continuation of U.S. application Ser. No.12/808,249, filed Jun. 15, 2010. U.S. application Ser. No. 12/808,249 isa 35 U.S.C. §371 national phase application of International ApplicationSerial No. PCT/GB2008/004208 (WO 2009/077766) filed Dec. 19, 2008,entitled “Pyrido[2,3-b]pyrazin-8-substituted Compounds and Their Use.”International Application Serial No. PCT/GB2008/004208 is anon-provisional application of U.S. provisional patent application No.61/015,019 filed Dec. 19, 2007, the contents of all of which areincorporated herein by reference in their entirety.

TECHNICAL FIELD

The present invention pertains generally to the field of therapeuticcompounds for treating proliferative disorders, cancer, etc., and morespecifically to certain pyrido[2,3-b]pyrazin-8-substituted compounds, asdescribed herein, which, inter alia, inhibit RAF (e.g., B-RAF) activity.The present invention also pertains to pharmaceutical compositionscomprising such compounds, and the use of such compounds andcompositions, both in vitro and in vivo, to inhibit RAF (e.g., BRAF)activity, to inhibit receptor tyrosine kinase (RTK) activity, to inhibitcell proliferation, and in the treatment of diseases and disorders thatare ameliorated by the inhibition of RAF, RTK, etc., proliferativedisorders such as cancer (e.g., colorectal cancer, melanoma), etc.

BACKGROUND

A number of patents and publications are cited herein in order to morefully describe and disclose the invention and the state of the art towhich the invention pertains. Each of these references is incorporatedherein by reference in its entirety into the present disclosure, to thesame extent as if each individual reference was specifically andindividually indicated to be incorporated by reference.

Throughout this specification, including the claims which follow, unlessthe context requires otherwise, the word “comprise,” and variations suchas “comprises” and “comprising,” will be understood to imply theinclusion of a stated integer or step or group of integers or steps butnot the exclusion of any other integer or step or group of integers orsteps.

It must be noted that, as used in the specification and the appendedclaims, the singular forms “a,” “an,” and “the” include plural referentsunless the context clearly dictates otherwise. Thus, for example,reference to “a pharmaceutical carrier” includes mixtures of two or moresuch carriers, and the like.

Ranges are often expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, another embodiment includes from the one particular valueand/or to the other particular value. Similarly, when values areexpressed as approximations, by the use of the antecedent “about,” itwill be understood that the particular value forms another embodiment.

This disclosure includes information that may be useful in understandingthe present invention. It is not an admission that any of theinformation provided herein is prior art or relevant to the presentlyclaimed invention, or that any publication specifically or implicitlyreferenced is prior art.

RAF, Proliferative Disorders, and Cancer

Mutations in genes that directly or indirectly control cell growth anddifferentiation are generally considered to be the main cause of cancer.Malignant tumors develop through a series of stepwise, progressivechanges that lead to the loss of growth control characteristic of cancercells, i.e., continuous unregulated proliferation, the ability to invadesurrounding tissues, and the ability to metastasize to different organsites. Carefully controlled in vitro studies have helped define thefactors that characterize the growth of normal and neoplastic cells andhave led to the identification of specific proteins that control cellgrowth and differentiation.

RAF is key downstream target for the ras GTPase and mediates theactivation of the MAP kinase cascade consisting of raf-MEK-ERK.Activated ERK is a kinase that subsequently targets a number of proteinsresponsible for mediating, amongst other things, the growth, survivaland transcriptional functions of the pathway. These include thetranscription factors ELK1, C-JUN, the Ets family (including Ets 1, 2,and 7), and the FOS family. The ras-raf-MEK-ERK signal transductionpathway is activated in response to many cell stimuli including growthfactors such as EGF, PDGF, KGF etc. Because the pathway is a majortarget for growth factor action, the activity of raf-MEK-ERK has beenfound to be upregulated in many factor dependent tumours. Theobservation that about 20% of all tumours have undergone an activatingmutation in one of the ras proteins indicates that the pathway is morebroadly important in tumorigenesis. There is growing evidence thatactivating mutations in other components of the pathway also occur inhuman tumours. This is true for RAF.

The RAF oncogene family includes three highly conserved genes termedA-RAF, B-RAF and C-RAF (also called Raf-1). RAF genes encode proteinkinases that are thought to play important regulatory roles in signaltransduction processes that regulate cell proliferation. RAF genes codefor highly conserved serine-threonine-specific protein kinases, whichare recruited to the plasma membrane following direct binding to the Rassmall Guanine-nucleotide binding proteins and this is the initiatingevent in RAF activation. RAF proteins are part of a signal transductionpathway believed to consist of receptor tyrosine kinases, p21 Ras, RAFprotein kinases, Mek1 (ERK activator or MAPKK) kinases and ERK (MAPK)kinases, which ultimately phosphorylate several cellular substrates,including transcription factors. Signaling through this pathway canmediate differentiation, proliferation or oncogenic transformation indifferent cellular contexts. Thus, RAF kinases are believed to play afundamental role in the normal cellular signal transduction pathway,coupling a multitude of growth factors to their net effect, cellularproliferation. Because RAF proteins are direct downstream effectors ofras protein function, therapies directed against RAF kinases arebelieved to be useful in treatment of ras-dependent tumors.

The RAF kinases are differentially regulated and expressed; C-RAF is themost thoroughly characterized and is expressed in all organs and in allcell lines that have been examined. A-RAF and B-RAF also appear to beubiquitous, but are most highly expressed in urogenital and braintissues, respectively. Because B-RAF is highly expressed in neuraltissues it was once thought to be limited to these tissues but it hassince been found to be more widely expressed. Although all RAF proteinscan bind to active Ras, B-raf is most strongly activated by oncogenicRas, and may be the primary target of oncogenic Ras in transformedcells.

Recent evidence indicates that mutational activation of B-RAF is foundin a number of different tumours including more than 65% of malignantmelanomas, more than 10% of colorectal cancers (Davies, H., et al.,2002, Nature, Vol. 417, pp. 949-954; Rajagopalan, H. et al., 2002,Nature, Vol. 418, p. 934), ovarian cancers (Singer, G., et al., 2003, J.Natl. Cancer Inst., Vol. 95, pp. 484-486) and papillary thyroid cancers(Brose, M., et al., 2002, Cancer Res., Vol. 62, pp. 6997-7000; Cohen,Y., et al., 2003, Invest. Ophthalmol. Vis. Sci., Vol. 44, pp.2876-2878). A range of different B-RAF mutations have been identified indifferent tumours with the most common being a V600E mutation in theso-called activation loop of the kinase domain (Davies, H., et al.,2002, Nature, Vol. 417, pp. 949-954).

Other mutations of B-RAF found associated with human cancers may notnecessarily activate B-RAF directly but do upregulate the activity ofthe ras-raf-MEK-ERK pathway by mechanisms which are not fully understoodbut may involve cross-talk with other RAF isoforms, such as A-RAF (Wan,P., et al., 2004, Cell, Vol. 116, pp. 855-867). In such cases,inhibition of RAF activity would remain a beneficial aim in cancertreatment.

In addition to link between B-RAF and certain cancers, there is asignificant amount of evidence to indicate a more broad inhibition ofRAF activity could be beneficial as an antitumour therapy. Blocking thepathway at the level of B-RAF would be effective at counteracting theupregulation of this pathway caused by tumourigenic ras mutations andalso in tumours responding to growth factor action via this pathway.Genetic evidence in Drosophila and C. elegans indicates that RAFhomologues are essential for ras dependent actions on differentiation(Dickson, B., et al., 1993, Nature, Vol. 360, pp. 600-603). Introductionof constitutively active MEK into NIH3T3 cells can have a transformingaction whilst expression of dominant negative MEK proteins can suppressthe tumourigenicity of ras transformed cell lines (Mansour, S. J., etal., 1994, Science, Vol. 265, pp. 966-970; Cowely, S., et al., 1994,Cell, Vol. 77, pp. 841-852). Expression of a dominant negative rafprotein has also been found to inhibit ras dependent signalling as hassuppression of raf expression using an antisense oligonucleotideconstruct (Koch, W., et al., 1991, Nature, Vol. 349, pp. 426-428;Bruder, T. T., et al., 1992, Genes and Development, Vol. 6, pp.545-556).

This and other evidence suggests that inhibition of RAF (e.g., B-RAF)activity would be beneficial in the treatment of cancer, and thatinhibition of RAF (e.g., B-RAF) activity could be particularlybeneficial in those cancers containing a constitutively activated B-rafmutation.

The raf-MEK-ERK pathway functions downstream of many receptors andstimuli indicating a broad role in regulation of cell function. For thisreason inhibitors of RAF may find utility in other disease conditionsthat are associated with upregulation of signalling via this pathway.The raf-MEK-ERK pathway is also an important component of the normalresponse of non-transformed cells to growth factor action. Thereforeinhibitors of RAF may be of use in diseases where there is inappropriateor excessive proliferation of normal tissues. These include, but are notlimited to glomerulonephritis and psoriasis. The cellular signallingpathway of which RAF is a part has also been implicated in inflammatorydisorders characterized by T-cell proliferation (T-cell activation andgrowth), such as tissue graft rejection, endotoxin shock, and glomerularnephritis.

RAF (e.g., B-RAF) has been shown to be a valid therapeutic target inhyperproliferative disorders such as cancer. Activated versions of RAF(e.g., B-RAF) are able to transform mammalian cells, allowing them totake on the characteristics of cancer cells and the growth of thesecells becomes dependent on the mutant RAF (e.g., B-RAF) protein.Inhibition of RAF (e.g., B-RAF) activity in human cancer cell lines thatexpress the mutant forms of RAF (e.g., B-RAF) blocks their growth andultimately induces their death.

Angiogenesis

Chronic proliferative diseases are often accompanied by profoundangiogenesis, which can contribute to or maintain an inflammatory and/orproliferative state, or which leads to tissue destruction through theinvasive proliferation of blood vessels. (Folkman, 1997, EXS, Vol. 79,pp. 1-81; Folkman, 1995, Nature Medicine, Vol. 1, pp. 27-31; Folkman andShing, 1992, J. Biol. Chem., Vol. 267, p. 10931.)

Angiogenesis is generally used to describe the development of new orreplacement blood vessels, or neovascularisation. It is a necessary andphysiological normal process by which the vasculature is established inthe embryo. Angiogenesis does not occur, in general, in most normaladult tissues, exceptions being sites of ovulation, menses and woundhealing. Many diseases, however, are characterized by persistent andunregulated angiogenesis. For instance, in arthritis, new capillaryblood vessels invade the joint and destroy cartilage (Colville-Nash andScott, 1992, Ann. Rhum. Dis., Vol. 51, p. 919). In diabetes (and in manydifferent eye diseases), new vessels invade the macula or retina orother ocular structures, and may cause blindness (Brooks et al., 1994,Cell, Vol. 79, p. 1157). The process of atherosclerosis has been linkedto angiogenesis (Kahlon et al., 1992, Can. J. Cardiol., Vol. 8, p. 60).Tumor growth and metastasis have been found to be angiogenesis-dependent(Folkman, 1992, Cancer Biol., Vol. 3, p. 65; Denekamp, 1993, Br. J.Rad., Vol. 66, p. 181; Fidler and Ellis, 1994, Cell, Vol. 79, p. 185).

The recognition of the involvement of angiogenesis in major diseases hasbeen accompanied by research to identify and develop inhibitors ofangiogenesis. These inhibitors are generally classified in response todiscrete targets in the angiogenesis cascade, such as activation ofendothelial cells by an angiogenic signal; synthesis and release ofdegradative enzymes; endothelial cell migration; proliferation ofendothelial cells; and formation of capillary tubules. Therefore,angiogenesis occurs in many stages and attempts are underway to discoverand develop compounds that work to block angiogenesis at these variousstages.

There are publications that teach that inhibitors of angiogenesis,working by diverse mechanisms, are beneficial in diseases such as cancerand metastasis (O'Reilly et al., 1994, Cell, Vol. 79, p. 315; Ingber etal., 1990, Nature, Vol. 348, p. 555), ocular diseases (Friedlander etal., 1995, Science, Vol. 270, p. 1500), arthritis (Peacock et al., 1992,J. Exp. Med., Vol. 175, p. 1135; Peacock et al., 1995, Cell. Immun.,Vol. 160, p. 178) and hemangioma (Taraboletti et al., 1995, J. Natl.Cancer Inst., Vol. 87, p. 293).

RTKs

Receptor tyrosine kinases (RTKs) are important in the transmission ofbiochemical signals across the plasma membrane of cells. Thesetransmembrane molecules characteristically consist of an extracellularligand-binding domain connected through a segment in the plasma membraneto an intracellular tyrosine kinase domain. Binding of ligand to thereceptor results in stimulation of the receptor-associated tyrosinekinase activity that leads to phosphorylation of tyrosine residues onboth the receptor and other intracellular proteins, leading to a varietyof cellular responses. To date, at least nineteen distinct RTKsubfamilies, defined by amino acid sequence homology, have beenidentified.

FGFR

The fibroblast growth factor (FGF) family of signaling polypeptidesregulates a diverse array of physiologic functions includingmitogenesis, wound healing, cell differentiation and angiogenesis, anddevelopment. Both normal and malignant cell growth as well asproliferation are affected by changes in local concentration of theseextracellular signaling molecules, which act as autocrine as well asparacrine factors. Autocrine FGF signaling may be particularly importantin the progression of steroid hormone-dependent cancers and to a hormoneindependent state (Powers et al., 2000, Endocr. Relat. Cancer, Vol. 7,pp. 165-197).

FGFs and their receptors are expressed at increased levels in severaltissues and cell lines and overexpression is believed to contribute tothe malignant phenotype. Furthermore, a number of oncogenes arehomologues of genes encoding growth factor receptors, and there is apotential for aberrant activation of FGF-dependent signaling in humanpancreatic cancer (Ozawa et al., 2001, Teratog. Carcinog. Mutagen., Vol.21, pp. 27-44).

The two prototypic members are acidic fibroblast growth factor (aFGF orFGF1) and basic fibroblast growth factors (bFGF or FGF2), and to date,at least twenty distinct FGF family members have been identified. Thecellular response to FGFs is transmitted via four types of high affinitytransmembrane tyrosine-kinase fibroblast growth factor receptorsnumbered 1 to 4 (FGFR-1 to FGFR-4). Upon ligand binding, the receptorsdimerize and auto- or trans-phosphorylate specific cytoplasmic tyrosineresidues to transmit an intracellular signal that ultimately reachesnuclear transcription factor effectors.

Disruption of the FGFR-1 pathway should affect tumor cell proliferationsince this kinase is activated in many tumor types in addition toproliferating endothelial cells. The over-expression and activation ofFGFR-1 in tumor-associated vasculature has suggested a role for thesemolecules in tumor angiogenesis.

FGFR-2 has high affinity for the acidic and/or basic fibroblast growthfactors, as well as the keratinocyte growth factor ligands. FGFR-2 alsopropagates the potent osteogenic effects of FGFs during osteoblastgrowth and differentiation. Mutations in FGFR-2, leading to complexfunctional alterations, were shown to induce abnormal ossification ofcranial sutures(craniosynostosis), implying a major role of FGFRsignaling in intramembranous bone formation. For example, in Apert (AP)syndrome, characterized by premature cranial suture ossification, mostcases are associated with point mutations engendering gain-of-functionin FGFR-2 (Lemonnier et al., 2001, J. Bone Miner. Res., Vol. 16, pp.832-845).

Several severe abnormalities in human skeletal development, includingApert, Crouzon, Jackson-Weiss, Beare-Stevenson cutis gyrata, andPfeiffer syndromes are associated with the occurrence of mutations inFGFR-2. Most, if not all, cases of Pfeiffer Syndrome (PS) are alsocaused by de novo mutation of the FGFR-2 gene (Meyers et al., 1996, Am.J. Hum. Genet., Vol. 58, pp. 491-498; Plomp et al., 1998, Am. J. Med.Genet., Vol. 75, 245-251), and it was recently shown that mutations inFGFR-2 break one of the cardinal rules governing ligand specificity.Namely, two mutant splice forms of fibroblast growth factor receptor,FGFR2c and FGFR2b, have acquired the ability to bind to and be activatedby atypical FGF ligands. This loss of ligand specificity leads toaberrant signaling and suggests that the severe phenotypes of thesedisease syndromes result from ectopic ligand-dependent activation ofFGFR-2 (Yu et al., 2000, Proc. Natl. Acad. Sci. U.S.A., Vol. 97, pp.14536-14541).

Activating mutations of the FGFR-3 receptor tyrosine kinase such aschromosomal translocations or point mutations produce deregulated,constitutively active, FGFR-3 receptors which have been involved inmultiple myeloma and in bladder and cervix carcinomas (Powers, C. J., etal., 2000, Endocr. Rel. Cancer, Vol. 7, p. 165). Accordingly, FGFR-3inhibition would be useful in the treatment of multiple myeloma, bladderand cervix carcinomas.

VEGFR

Vascular endothelial growth factor (VEGF), a polypeptide, is mitogenicfor endothelial cells in vitro and stimulates angiogenic responses invivo. VEGF has also been linked to inappropriate angiogenesis (Pinedo,H. M., et al., 2000, The Oncologist, Vol. 5 (90001), pp. 1-2). VEGFR(s)are protein tyrosine kinases (PTKs). PTKs catalyze the phosphorylationof specific tyrosyl residues in proteins involved in the regulation ofcell growth and differentiation. (Wilks, A. F., 1990, Progress in GrowthFactor Research, Vol. 2, pp. 97-111; Courtneidge, S. A., 1993, Dev.Supp.l, pp. 57-64; Cooper, J. A., 1994, Semin. Cell Biol., Vol. 5(6),pp. 377-387; Paulson, R. F., 1995, Semin. Immunol., Vol. 7(4), pp.267-277; Chan, A. C., 1996, Curr. Opin. Immunol., Vol. 8(3), pp.394-401).

Three PTK receptors for VEGF have been identified: VEGFR-1 (Flt-1),VEGFR-2 (Flk-1 or KDR), and VEGFR-3 (Flt-4). These receptors areinvolved in angiogenesis and participate in signal transduction(Mustonen, T., et al., 1995, J. Cell Biol., Vol. 129, pp. 895-898).

Of particular interest is VEGFR-2, which is a transmembrane receptor PTKexpressed primarily in endothelial cells. Activation of VEGFR-2 by VEGFis a critical step in the signal transduction pathway that initiatestumour angiogenesis. VEGF expression may be constitutive to tumour cellsand can also be upregulated in response to certain stimuli.

One such stimuli is hypoxia, where VEGF expression is upregulated inboth tumour and associated host tissues. The VEGF ligand activatesVEGFR-2 by binding with its extracellular VEGF binding site. This leadsto receptor dimerization of VEGFRs and autophosphorylation of tyrosineresidues at the intracellular kinase domain of VEGFR-2. The kinasedomain operates to transfer a phosphate from ATP to the tyrosineresidues, thus providing binding sites for signalling proteinsdownstream of VEGFR-2 leading ultimately to initiation of angiogenesis(McMahon, G., 2000, The Oncologist, Vol. 5(90001), pp. 3-10).

Inhibition at the kinase domain binding site of VEGFR-2 would blockphosphorylation of tyrosine residues and serve to disrupt initiation ofangiogenesis.

TIE

Angiopoieten 1 (Ang1), a ligand for the endothelium-specific receptortyrosine kinase TIE-2 is a novel angiogenic factor (Davis et al., 1996,Cell, Vol. 87, pp. 1161-1169; Partanen et al., 1992, Mol. Cell Biol.,Vol. 12, pp. 1698-1707; U.S. Pat. Nos. 5,521,073; 5,879,672; 5,877,020;and 6,030,831). The acronym TIE represents “tyrosine kinase containingIg and EGF homology domains”. TIE is used to identify a class ofreceptor tyrosine kinases, which are exclusively expressed in vascularendothelial cells and early hemopoietic cells. Typically, TIE receptorkinases are characterized by the presence of an EGF-like domain and animmunoglobulin (IG) like domain, which consists of extracellular foldingunits, stabilized by intra-chain disulfide bonds (Partanen et al., 1999,Curr. Topics Microbiol. Immunol., Vol. 237, pp. 159-172). Unlike VEGF,which functions during the early stages of vascular development, Ang1and its receptor TIE-2 function in the later stages of vasculardevelopment, i.e., during vascular remodelling (remodelling refers toformation of a vascular lumen) and maturation (Yancopoulos et al., 1998,Cell, Vol. 93, pp. 661-664; Peters, K. G., 1998, Circ. Res., Vol. 83(3),pp. 342-343; Suri et al., 1996, Cell, Vol. 87, pp. 1171-1180).

Consequently, inhibition of TIE-2 would be expected to serve to disruptremodelling and maturation of new vasculature initiated by angiogenesisthereby disrupting the angiogenic process.

Eph

The largest subfamily of receptor tyrosine kinases (RTKs), the Ephfamily, and their ligands (ephrins), play important roles in physiologicand pathologic vascular processes. Both the Ephs (receptors) and ephrins(ligands) are divided into two groups, A and B subfamilies (EphNomenclature Committee, 1997). The binding of ephrin ligands to Ephreceptors is dependent on cell-cell interactions. The interactions ofephrins and Ephs have recently been shown to function via bi-directionalsignalling. The ephrins binding to Eph receptors initiatephosphorylation at specific tyrosine residues in the cytoplasmic domainof the Eph receptors. In response to Eph receptor binding, the ephrinligand also undergoes tyrosine phosphorylation, so-called ‘reverse’signalling (Holland, S. J., et al., 1996, Nature, Vol. 383, pp. 722-725;Bruckner et al., 1997, Science, Vol. 275, pp. 1640-1643).

Eph RTKs and their ephrin ligands play important roles in embryonicvascular development. Disruption of specific Eph receptors and ligands(including ephrin-B2) leads to defective vessel remodelling,organisation, and sprouting resulting in embryonic death (Wang, H. U.,et al., 1998, Cell, Vol. 93, pp. 741-753; Adams, R. H., et al., 1999,Genes Dev, Vol. 13, pp. 295-306; Gale and Yancopoulos, 1999, Genes Dev,Vol. 13, pp. 1055-1066; Helbling, P. M., et al., 2000, Development, Vol.127, pp. 269-278). Coordinated expression of the Eph/ephrin systemdetermines the phenotype of embryonic vascular structures: ephrin-B2 ispresent on arterial endothelial cells (ECs), whereas EphB4 is present onvenous ECs (Gale and Yancopoulos, 1999, Genes Dev, Vol. 13, pp.1055-1066; Shin, D., et al., 2001, Dev Biol, Vol. 230, pp. 139-150).Recently, specific Ephs and ephrins have been implicated in tumourgrowth and angiogenesis.

The Ephs and ephrins have been found to be overexpressed in many humantumours. In particular, the role of EphB2 has been identified in smallcell lung carcinoma (Tang, X. X., et al., 1999, Clin Cancer Res, Vol. 5,pp. 455-460), human neuroblastomas (Tang, X. X., et al., 1999, ClinCancer Res, Vol. 5, pp. 1491-1496) and colorectal cancers (Liu, W., etal., 2004, Brit. J. Canc., Vol. 90, pp. 1620-1626), and higherexpression levels of Ephs and ephrins, including EphB2, have been foundto correlate with more aggressive and metastatic tumours (Nakamoto, M.and Bergemann, A. D., 2002, Microsc. Res Tech, Vol. 59, pp. 58-67).

Consequently, inhibition of EphB2 would be expected to serve to disruptangiogenesis, and in particular in certain tumours where over-expressionoccurs.

The inventors have discovered compounds that, e.g., inhibit RAF (e.g.,B-RAF) activity and/or are useful in the treatment of, e.g.,proliferative disorders, cancer, etc.

SUMMARY OF THE INVENTION

One aspect of the invention pertains to certainpyrido[2,3-b]pyrazin-8-substituted compounds (referred to herein as“PDP8 compounds”), as described herein.

Another aspect of the invention pertains to a composition (e.g., apharmaceutical composition) comprising a PDP8 compound, as describedherein, and a pharmaceutically acceptable carrier or diluent.

Another aspect of the invention pertains to method of preparing acomposition (e.g., a pharmaceutical composition) comprising the step ofadmixing a PDP8 compound, as described herein, and a pharmaceuticallyacceptable carrier or diluent.

Another aspect of the present invention pertains to a method ofinhibiting RAF (e.g., B-RAF) activity in a cell, in vitro or in vivo,comprising contacting the cell with an effective amount of a PDP8compound, as described herein.

Another aspect of the present invention pertains to a method ofinhibiting receptor tyrosine kinase (RTK) activity, such as FGFR, Tie,VEGFR and/or Eph activity, for example, FGFR-1, FGFR-2, FGFR-3, Tie2,VEGFR-2 and/or EphB2 activity, in a cell, in vitro or in vivo,comprising contacting the cell with an effective amount of a PDP8compound, as described herein.

Another aspect of the present invention pertains to a method ofregulating (e.g., inhibiting) cell proliferation (e.g., proliferation ofa cell), inhibiting cell cycle progression, promoting apoptosis, or acombination of one or more these, in vitro or in vivo, comprisingcontacting a cell with an effective amount of a PDP8 compound, asdescribed herein.

Another aspect of the present invention pertains to a method oftreatment comprising administering to a subject in need of treatment atherapeutically-effective amount of a PDP8 compound, as describedherein, preferably in the form of a pharmaceutical composition.

Another aspect of the present invention pertains to a PDP8 compound, asdescribed herein, for use in a method of treatment of the human oranimal body by therapy.

Another aspect of the present invention pertains to a PDP8 compound, asdescribed herein, for the use in a method of treatment of the human oranimal body by therapy wherein said compound is used in combination withother pharmaceutically active substances

Another aspect of the present invention pertains to use of a PDP8compound, as described herein, in the manufacture of a medicament foruse in treatment.

In one embodiment, the treatment is treatment of a disease or disorder(e.g., cancer) that is characterised by the up-regulation and/oractivation of RAF (e.g., B-RAF), and/or is ameliorated by the inhibitionof RAF (e.g., B-RAF).

In one embodiment, the treatment is treatment of a disease or disorder(e.g., cancer) that is characterised by the up-regulation and/oractivation of a receptor tyrosine kinase (RTK), and/or is ameliorated bythe inhibition of a receptor tyrosine kinase (RTK). Examples of RTKsinclude FGFR, Tie, VEGFR and/or Eph, for example, FGFR-1, FGFR-2,FGFR-3, Tie2, VEGFR-2 and/or EphB2.

In one embodiment, the treatment is treatment of a disease or disorderthat is characterised by inappropriate, excessive, and/or undesirableangiogenesis.

In one embodiment, the treatment is treatment of a proliferativedisorder.

In one embodiment, the treatment is treatment of cancer.

In one embodiment, the treatment is treatment of melanoma.

In one embodiment, the treatment is treatment of colorectal cancer.

Another aspect of the present invention pertains to a kit comprising (a)a PDP8 compound, as described herein, preferably provided as apharmaceutical composition and in a suitable container and/or withsuitable packaging; and (b) instructions for use, for example, writteninstructions on how to administer the compound.

Another aspect of the present invention pertains to a PDP8 compoundobtainable by a method of synthesis as described herein, or a methodcomprising a method of synthesis as described herein.

Another aspect of the present invention pertains to a PDP8 compoundobtained by a method of synthesis as described herein, or a methodcomprising a method of synthesis as described herein.

Another aspect of the present invention pertains to novel intermediates,as described herein, which are suitable for use in the methods ofsynthesis described herein.

Another aspect of the present invention pertains to the use of suchnovel intermediates, as described herein, in the methods of synthesisdescribed herein.

As will be appreciated by one of skill in the art, features andpreferred embodiments of one aspect of the invention will also pertainto other aspect of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 1 (AA-018) (non-established) (5 mg/kg/day)(intraperitoneally).

FIG. 2 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 2 (AA-018) (non-established) (10mg/kg/day) (intraperitoneally).

FIG. 3 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 3 (AA-019) (non-established) (5 mg/kg/day)(intraperitoneally).

FIG. 4 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 4 (AA-019) (non-established) (10mg/kg/day) (intraperitoneally).

FIG. 5 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 5 (AA-019) (non-established) (15mg/kg/day) (orally).

FIG. 6 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 6 (AA-019) (established) (10/5 mg/kg/day)(intraperitoneally).

FIG. 7 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 7 (AA-019) (established) (15 mg/kg/day)(orally).

FIG. 8 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 8 (AA-062) (established) (50 mg/kg/day)(orally).

FIG. 9 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 9 (AA-067) (established) (10 mg/kg/day)(orally).

FIG. 10 is a graph of relative tumour volume as a function of days frominoculation for In Vivo Study 10 (AA-017) (established) (20 mg/kg/day)(orally).

DETAILED DESCRIPTION OF THE INVENTION Compounds

One aspect of the present invention pertains to compounds selected fromcompounds of the following formula and pharmaceutically acceptablesalts, hydrates, and solvates thereof (for convenience, collectivelyreferred to herein as “pyrido[2,3-b]pyrazin-8-substituted compounds” and“PDP8 compounds”):

wherein:—R^(Q1) is independently —H, —R¹, —R^(1X), —Cl, —OH, —OR¹, —OR^(1X),—SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB);wherein:

-   -   each —R¹ is independently saturated aliphatic C₁₋₆alkyl, and is        unsubstituted or substituted, for example, with one or more        groups selected from —OH, —OR¹¹, —NH₂, —NHR¹¹, and —NR¹¹ ₂,        wherein each —R¹¹ is independently saturated aliphatic        C₁₋₃alkyl;    -   each —R^(1X) is independently saturated aliphatic C₁₋₄alkyl        substituted with one or more groups selected from —F, —Cl, —Br,        and —I; and    -   —NR^(RA)R^(RB) is independently azetidino, pyrrolidino,        piperidino, piperazino, morpholino, azepino, or diazepino, and        is optionally substituted with one or more groups selected from        saturated aliphatic C₁₋₄alkyl;        —R^(Q2) is independently —H, —R^(2X), —Cl, —OH, —OR², —OR^(2X),        —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD);        wherein:    -   each —R² is independently saturated aliphatic C₁₋₆alkyl, and is        unsubstituted or substituted, for example, with one or more        groups selected from —OH, —OR²², —NH₂, —NHR²², and —NR²² ₂,        wherein each —R²² is independently saturated aliphatic        C₁₋₃alkyl;    -   each —R^(2X) is independently saturated aliphatic C₁₋₄alkyl        substituted with one or more groups selected from —F, —Cl, —Br,        and —I; and    -   —NR^(RC)R^(RD) is independently azetidino, pyrrolidino,        piperidino, piperazino, morpholino, azepino, or diazepino, and        is optionally substituted with one or more groups selected from        saturated aliphatic C₁₋₄alkyl;        —X— is independently —O—, —S—, —S(═O)—, or —S(═O)₂—;        -M- is independently selected from:

wherein:

-   -   each n is independently 0, 1 or 2; and    -   each R^(PH1) is independently —F, —Cl, —Br, —I, —R³, —R^(3Y),        —CF₃, —OH, —OR³, —OCF₃, —NH₂, —NHR³, —NR³ ₂, —CN, —SH, or —SR³;    -   wherein each —R³ is independently saturated aliphatic C₁₋₄alkyl,        and each —R^(3Y) is independently aliphatic C₂₋₆alkenyl or        aliphatic C₂₋₆alkynyl;        J-L- is independently selected from:    -   J-NR^(N1)—C(═Y)—NR^(N1)—,    -   J-CH₂—NR^(N1)—C(═Y)—NR^(N1)—,    -   J-NR^(N1)—C(═Y)—NR^(N1)—CH₂—,    -   J-NR^(N1)—C(═Y)—,    -   J-CH₂—NR^(N1)—C(═Y)—,    -   J-NR^(N1)—C(═Y)—CH₂—,    -   J-CH₂—NR^(N1)—C(═Y)—CH₂—,    -   J-CH₂—CH₂—NR^(N1)—C(═Y)—,    -   J-NR^(N1)—C(═Y)—CH₂—CH₂—,    -   J-NR^(N1)—C(═Y)—CH₂—NR^(N1)—,    -   J-NR^(N1)—CH₂—NR^(N1)—C(═Y)—,    -   J-C(═Y)—NR^(N1)—,    -   J-CH₂—C(═Y)—NR^(N1)—,    -   J-C(═Y)—NR^(N1)—CH₂—,    -   J-CH₂—C(═Y)—NR^(N1)—CH₂—,    -   J-CH₂—CH₂—C(═Y)—NR^(N1)—,    -   J-C(═Y)—NR^(N1)—CH₂—CH₂—,    -   J-NR^(N1)—CH₂—C(═Y)—NR^(N1)—,    -   J-C(═Y)—NR^(N1)—CH₂—NR^(N1)—,    -   J-C(═Y)—CH₂—NR^(N1)—,    -   J-C(═Y)—CH₂—NR^(N1)—CH₂—,    -   J-C(═Y)—CH₂—CH₂—NR^(N1)—,    -   J-CH₂—C(═Y)—CH₂—NR^(N1)—,    -   J-NR^(N1)—CH₂—C(═Y)—,    -   J-NR^(N1)—CH₂—C(═Y)—CH₂—,    -   J-NR^(N1)—CH₂—CH₂—C(═Y)—,    -   J-CH₂—NR^(N1)—CH₂—C(═Y)—,    -   J-NR^(N1)—S(═O)₂—NR^(N1)—,    -   J-NR^(N1)—S(═O)₂—NR^(N1)—CH₂—,    -   J-CH₂—NR^(N1)—S(═O)₂—NR^(N1)—,    -   J-NR^(N1)—S(═O)₂—,    -   J-NR^(N1)—S(═O)₂—CH₂—,    -   J-CH₂—NR^(N1)—S(═O)₂—,    -   J-CH₂—NR^(N1)—S(═O)₂—CH₂—,    -   J-CH₂—CH₂—NR^(N1)—S(═O)₂—,    -   J-NR^(N1)—S(═O)₂—CH₂—CH₂—,    -   J-NR^(N1)—S(═O)₂—CH₂—NR^(N1)—,    -   J-NR^(N1)—CH₂—NR^(N1)—S(═O)₂—,    -   J-S(═O)₂—NR^(N1)—,    -   J-S(═O)₂—NR^(N1)—CH₂—,    -   J-CH₂—S(═O)₂—NR^(N1)—,    -   J-CH₂—S(═O)₂—NR^(N1)—CH₂—,    -   J-CH₂—CH₂—S(═O)₂—NR^(N1)—,    -   J-S(═O)₂—NR^(N1)—CH₂—CH₂—,    -   J-S(═O)₂—NR^(N1)—CH₂—NR^(N1)—, and    -   J-NR^(N1)—CH₂—S(═O)₂—NR^(N1)—;    -   wherein:    -   each —R^(N1) is independently —H or saturated aliphatic        C₁₋₄alkyl; and    -   each ═Y is independently ═O or ═S; and        -J is independently phenyl or C₅₋₆heteroaryl, and is optionally        substituted, for example, with one or more substituents selected        from:    -   —F, —Cl, —Br, —I, —CF₃, —OCF₃,    -   R⁴, —R^(4S), —R^(4A), —R^(4B), —R^(4C), -L^(4C), —Ar, -L⁴-Ar,    -   —OH, —OR⁴, -L⁴-OH, -L⁴-OR⁴, —O-L⁴-OH, —O-L⁴-OR⁴,    -   —OR^(4C), —O-L⁴-R^(4C), —OAr, —O-L⁴-Ar,    -   —SH, —SR⁴, —CN, —NO₂,    -   —NH₂, —NHR^(4SS), —R^(N),    -   -L⁴-NH₂, -L⁴-NHR^(4SS), -L⁴-R^(N),    -   —O-L⁴-NH₂, —O-L⁴-NHR^(4SS), —O-L⁴-R^(N),    -   —NH-L⁴-NH₂, —NH-L⁴-NHR^(4SS), —NH-L⁴-R^(N),    -   —NR⁴-L⁴-NH₂, —NR⁴-L⁴-NHR^(4SS), —NR⁴-L⁴-R^(N),        wherein:    -   each —R⁴ is independently saturated aliphatic C₁₋₆alkyl;    -   each —R^(4S) is independently saturated aliphatic C₁₋₆alkyl        substituted with one or more groups selected from —OH,        —OR^(4SS), —C(═O)OH, —C(═O)OR^(4SS), —NH₂, —NHR^(4SS),        —N(R^(4SS))₂, —R^(N), —C(═O)NH₂, —C(═O)NHR^(4SS),        —C(═O)N(R^(4SS))₂, and —C(═O)R^(N);    -   each —R^(4A) is independently aliphatic C₂₋₆alkenyl;    -   each —R^(4B) is independently aliphatic C₂₋₆alkynyl;    -   each —R^(4C) is independently optionally substituted saturated        C₃₋₆cycloalkyl, for example, saturated C₃₋₆cycloalkyl optionally        substituted with one or more substituents selected from —F, —R⁵,        —OH, —OR⁵, —CF₃, and —OCF₃,    -   each -L⁴- is independently saturated aliphatic C₁₋₄alkylene;    -   each —Ar is optionally substituted phenyl or C₅₋₆heteroaryl, for        example, phenyl or C₅₋₆heteroaryl optionally substituted with        one or more substituents selected from —F, —Cl, —Br, —I, —R⁵,        —OH, —OR⁵, —CF₃, —OCF₃, and —S(═O)₂R⁵;    -   each —R^(4SS) is independently saturated aliphatic C₁₋₄alkyl;    -   each —R^(N) is independently azetidino, pyrrolidino, piperidino,        piperazino, morpholino, azepino, or diazepino, and is optionally        substituted with one or more groups selected from saturated        aliphatic C₁₋₄alkyl; and    -   each —R⁵ is independently saturated aliphatic C₁₋₄alkyl.

In one embodiment, the compound is selected from compounds of thefollowing formula, and pharmaceutically acceptable salts, hydrates, andsolvates thereof:

wherein:

-   -   —R^(Q1) is independently —H, —R¹, —Cl, —OH, —OR¹, —SH, —SR¹,        —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB);    -   wherein:    -   each —R¹ is independently saturated aliphatic C₁₋₆alkyl, and is        unsubstituted or substituted, for example, with one or more        groups selected from —OH, —OR¹¹, —NH₂, —NHR¹¹, and —NR¹¹ ₂,        wherein each —R¹¹ is independently saturated aliphatic        C₁₋₃alkyl; and    -   —NR^(RA)R^(RB) is independently piperidino, piperazino, or        morpholino, and is optionally substituted with one or more        groups selected from saturated aliphatic C₁₋₄alkyl;    -   —R^(Q2) is independently —H, —R², —Cl, —OH, —OR², —SH, —SR²,        —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD);    -   wherein:    -   each —R² is independently saturated aliphatic C₁₋₆alkyl, and is        unsubstituted or substituted, for example, with one or more        groups selected from —OH, —OR²², —NH₂, —NHR²², and —NR²² ₂,        wherein each —R²² is independently saturated aliphatic        C₁₋₃alkyl; and    -   —NR^(RC)R^(RD) is independently piperidino, piperazino, or        morpholino, and is optionally substituted with one or more        groups selected from saturated aliphatic C₁₋₄alkyl;    -   —X— is independently —O— or —S—;    -   -M- is independently selected from:

-   -   wherein:        -   each n is independently 0, 1 or 2; and        -   each R^(PH1) is independently —F, —Cl, —Br, —I, —R³, —OH,            —OR³, —SH, or —SR³;        -   wherein each —R³ is independently saturated aliphatic            C₁₋₄alkyl;    -   -L- is independently selected from:

-   -   wherein:        -   each —R^(N1) is independently —H or saturated aliphatic            C₁₋₄alkyl; and    -   -J is independently phenyl or C₅₋₆heteroaryl, and is optionally        substituted, for example, with one or more substituents selected        from —F, —Cl, —Br, —I, —R⁴, —OH, —OR⁴, —CF₃, —OCF₃, and -Ph,        wherein each —R⁴ is independently saturated aliphatic C₁₋₄alkyl;        and each -Ph denotes optionally substituted phenyl, for example,        phenyl optionally substituted with one or more substituents        selected from —F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵, —CF₃, —OCF₃,        wherein each —R⁵ is independently saturated aliphatic C₁₋₄alkyl.

The Group —R^(Q1)

In one embodiment, —R^(Q1) is independently —H, —R¹, —R^(1X), —Cl, —OH,—OR¹, —OR^(1X), —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB).

-   -   In one embodiment, —R^(Q1) is independently —R¹, —R^(1X), —Cl,        —OH, —OR¹, —OR^(1X), —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or        —NR^(RA)R^(RB).    -   In one embodiment, —R^(Q1) is independently —H, —R¹, —R^(1X),        —Cl, —OR¹, —OR^(1X), —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or        —NR^(RA)R^(RB).    -   In one embodiment, —R^(Q1) is independently —R¹, —R^(1X), —Cl,        —OR¹, —OR^(1X), —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or        —NR^(RA)R^(RB).

In one embodiment, —R^(Q1) is independently —H, —R¹, —Cl, —OH, —OR¹,—SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB).

-   -   In one embodiment, —R^(Q1) is independently —R¹, —Cl, —OH, —OR¹,        —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB).    -   In one embodiment, —R^(Q1) is independently —H, —R¹, —Cl, —OR¹,        —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB).    -   In one embodiment, —R^(Q1) is independently —R¹, —Cl, —OR¹, —SH,        —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB).

In one embodiment, —R^(Q1) is independently —H, —OH, -Me, —CF₃, —CH₂Br,—NH₂, —NHMe, —NMe₂, morpholino, or piperazino, or N-methyl-piperazino.

-   -   In one embodiment, —R^(Q1) is independently —OH, -Me, —CF₃,        —CH₂Br, —NH₂, —NHMe, —NMe₂, morpholino, or piperazino, or        N-methyl-piperazino.    -   In one embodiment, —R^(Q1) is independently —H, -Me, —CF₃,        —CH₂Br, —NH₂, —NHMe, —NMe₂, morpholino, or piperazino, or        N-methyl-piperazino.    -   In one embodiment, —R^(Q1) is independently -Me, —CF₃, —CH₂Br,        —NH₂, —NHMe, —NMe₂, morpholino, or piperazino, or        N-methyl-piperazino.

In one embodiment, —R^(Q1) is independently —H, —OH, -Me, —NH₂, —NHMe,morpholino, or piperazino, or N-methyl-piperazino.

-   -   In one embodiment, —R^(Q1) is independently —OH, -Me, —NH₂,        —NHMe, morpholino, or piperazino, or N-methyl-piperazino.    -   In one embodiment, —R^(Q1) is independently —H, -Me, —NH₂,        —NHMe, morpholino, or piperazino, or N-methyl-piperazino.    -   In one embodiment, —R^(Q1) is independently -Me, —NH₂, —NHMe,        morpholino, or piperazino, or N-methyl-piperazino.

In one embodiment, —R^(Q1) is —OH. In this case, tautomerisation ispossible, and the two equivalent tautomers are shown below.

The Group —R^(Q2)

In one embodiment, —R^(Q2) is independently —H, —R², —R^(2X), —Cl, —OH,—OR², —OR^(2X), —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).

-   -   In one embodiment, —R^(Q2) is independently —R², —R^(2X), —Cl,        —OH, —OR², —OR^(2X), —SH, —SR², —NH₂, —NHR², —NR² ₂, or        —NR^(RC)R^(RD).    -   In one embodiment, —R^(Q2) is independently —H, —R², —R^(2X),        —Cl, —OR², —OR^(2X), —SH, —SR², —NH₂, —NHR², —NR² ₂, or        —NR^(RC)R^(RD).    -   In one embodiment, —R^(Q2) is independently —R², —R^(2X), —Cl,        —OR², —OR^(2X), —SH, —SR², —NH₂, —NHR², —NR² ₂, or        —NR^(RC)R^(RD).

In one embodiment, —R^(Q2) is independently —H, —R², —Cl, —OH, —OR²,—SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).

-   -   In one embodiment, —R^(Q2) is independently —R², —Cl, —OH, —OR²,        —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).    -   In one embodiment, —R^(Q2) is independently —H, —R², —Cl, —OR²,        —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).    -   In one embodiment, —R^(Q2) is independently —R², —Cl, —OR², —SH,        —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).

In one embodiment, —R^(Q2) is independently —H, —OH, -Me, —CF₃, —CH₂Br,—NH₂, —NHMe, —NMe₂, morpholino, or piperazino, or N-methyl-piperazino.

-   -   In one embodiment, —R^(Q2) is independently —OH, -Me, —CF₃,        —CH₂Br, —NH₂, —NHMe, —NMe₂, morpholino, or piperazino, or        N-methyl-piperazino.    -   In one embodiment, —R^(Q2) is independently —H, -Me, —CF₃,        —CH₂Br, —NH₂, —NHMe, —NMe₂, morpholino, or piperazino, or        N-methyl-piperazino.    -   In one embodiment, —R^(Q2) is independently -Me, —CF₃, —CH₂Br,        —NH₂, —NHMe, —NMe₂, morpholino, or piperazino, or        N-methyl-piperazino.

In one embodiment, —R^(Q2) is independently —H, —OH, -Me, —NH₂, —NHMe,morpholino, or piperazino, or N-methyl-piperazino.

-   -   In one embodiment, —R^(Q2) is independently —OH, -Me, —NH₂,        —NHMe, morpholino, or piperazino, or N-methyl-piperazino.    -   In one embodiment, —R^(Q2) is independently —H, -Me, —NH₂,        —NHMe, morpholino, or piperazino, or N-methyl-piperazino.    -   In one embodiment, —R^(Q2) is independently -Me, —NH₂, —NHMe,        morpholino, or piperazino, or N-methyl-piperazino.

In one embodiment, —R^(Q2) is —OH. In this case, tautomerisation ispossible, and the two equivalent tautomers are shown below.

Some Combinations of the Groups —R^(Q1) and —R^(Q2): Both are not —H

In one embodiment:either:

-   -   —R^(Q1) is independently —H, —R¹, —R^(1X), —Cl, —OH, —OR¹,        —OR^(1X), —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —R², —R^(2X), —Cl, —OH, —OR², —OR^(2X),        —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD);        or:    -   —R^(Q1) is independently —R¹, —R^(1X), —Cl, —OH, —OR¹, —OR^(1X),        —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —H, —R², —R^(2X), —Cl, —OH, —OR²,        —OR^(2X), —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:        either:    -   —R^(Q1) is independently —H, —OH, -Me, —CF₃, —CH₂Br, —NH₂,        —NHMe, —NMe₂, morpholino, or piperazino, or N-methyl-piperazino;        and    -   —R^(Q2) is independently —OH, -Me, —CF₃, —CH₂Br, —NH₂, —NHMe,        —NMe₂, morpholino, or piperazino, or N-methyl-piperazino;        or:    -   —R^(Q1) is independently —OH, -Me, —CF₃, —CH₂Br, —NH₂, —NHMe,        —NMe₂, morpholino, or piperazino, or N-methyl-piperazino; and    -   —R^(Q2) is independently —H, —OH, -Me, —CF₃, —CH₂Br, —NH₂,        —NHMe, —NMe₂, morpholino, or piperazino, or N-methyl-piperazino.

Some Combinations of the Groups —R^(Q1) and —R^(Q2): Exactly One is —OH

In one embodiment:either:

-   -   —R^(Q1) is independently —OH; and    -   —R^(Q2) is independently —H, —R², —R^(2X), —Cl, —OR², —OR^(2X),        —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD);        or:    -   —R^(Q1) is independently —H, —R¹, —R^(1X), —Cl, —OR¹, —OR^(1X),        —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —OH.        In one embodiment:    -   —R^(Q1) is independently —OH; and    -   —R^(Q2) is independently —H, —R², —R^(2X), —Cl, —OR², —OR^(2X),        —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:    -   —R^(Q1) is independently —H, —R¹, —R^(1X), —Cl, —OR¹, —OR^(1X),        —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —OH.        In one embodiment:        either:    -   —R^(Q1) is independently —OH; and    -   —R^(Q2) is independently —H, -Me, —CF₃, —CH₂Br, —NH₂, —NHMe,        —NMe₂, morpholino, or piperazino, or N-methyl-piperazino;        or:    -   —R^(Q1) is independently —H, -Me, —CF₃, —CH₂Br, —NH₂, —NHMe,        —NMe₂, morpholino, or piperazino, or N-methyl-piperazino; and    -   —R^(Q2) is independently —OH.        In one embodiment:    -   —R^(Q1) is independently —OH; and    -   —R^(Q2) is independently —H, -Me, —CF₃, —CH₂Br, —NH₂, —NHMe,        —NMe₂, morpholino, or piperazino, or N-methyl-piperazino.        In one embodiment:    -   —R^(Q1) is independently —H, -Me, —CF₃, —CH₂Br, —NH₂, —NHMe,        —NMe₂, morpholino, or piperazino, or N-methyl-piperazino; and    -   —R^(Q2) is independently —OH.        In one embodiment:        either:    -   —R^(Q1) is —OH, and    -   —R^(Q2) is independently —H, -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino.        or:    -   —R^(Q1) is independently —H, -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino, and    -   —R^(Q2) is —OH.        In one embodiment:    -   —R^(Q1) is —OH, and    -   —R^(Q2) is independently —H, -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino.        In one embodiment,    -   —R^(Q1) is independently —H, -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino, and    -   —R^(Q2) is —OH.        In one embodiment:        either:    -   —R^(Q1) is -Me or —NH², and    -   —R^(Q2) is —OH;        or:    -   —R^(Q1) is —OH, and    -   —R^(Q2) is -Me or —NH².        In one embodiment:    -   —R^(Q1) is -Me or —NH², and    -   —R^(Q2) is —OH.        In one embodiment:    -   —R^(Q1) is —OH, and    -   —R^(Q2) is -Me or —NH².        In one embodiment:        either:    -   —R^(Q1) is —OH, and    -   —R^(Q2) is —H;        or:    -   —R^(Q1) is —H, and    -   —R^(Q2) is —OH.        In one embodiment:    -   —R^(Q1) is —OH, and    -   —R^(Q2) is —H.        In one embodiment:    -   —R^(Q1) is —H, and    -   —R^(Q2) is —OH.

Some Combinations of the Groups —R^(Q1) and —R^(Q2): Both are —OH

In one embodiment:

-   -   —R^(Q1) is —OH and    -   —R^(Q2) is —OH.

In this case, tautomerisation is possible, and the two equivalenttautomers are shown below.

Some Combinations of the Groups —R^(Q1) and —R^(Q2): Neither is —OH

In one embodiment:

-   -   —R^(Q1) is independently —H, —R¹, —R^(1X), —Cl, —OR¹, —OR^(1X),        —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —H, —R², —R^(2X), —Cl, —OR², —OR^(2X),        —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:    -   —R^(Q1) is independently —H, —R¹, —Cl, —OR¹, —SH, —SR¹, —NH₂,        —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —H, —R², —Cl, —OR², —SH, —SR², —NH₂,        —NHR², —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:    -   —R^(Q1) is independently —H, -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino; and    -   —R^(Q2) is independently —H, -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino.        In one embodiment:    -   —R^(Q1) is independently —H; and    -   —R^(Q2) is independently —H, -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino.

Some Combinations of the Groups —R^(Q1) and —R^(Q2): Neither is —OH andBoth are not —H

In one embodiment:either:

-   -   —R^(Q1) is independently —H, —R¹, —R^(1X), —Cl, —OR¹, —OR^(1X),        —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —R², —R^(2X), —Cl, —OR², —OR^(2X), —SH,        —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD);        or:    -   —R^(Q1) is independently —R¹, —R^(1X), —Cl, —OR¹, —OR^(1X), —SH,        —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —H, —R², —R^(2X), —Cl, —OR², —OR^(2X),        —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:    -   —R^(Q1) is independently —H, —R¹, —R^(1X), —Cl, —OR¹, —OR^(1X),        —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —R², —R^(2X), —Cl, —OR², —OR^(2X), —SH,        —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:    -   —R^(Q1) is independently —R¹, —R^(1X), —Cl, —OR¹, —OR^(1X), —SH,        —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —H, —R², —R^(2X), —Cl, —OR², —OR^(2X),        —SH, —SR², —NH₂, —NHR², —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:        either:    -   —R^(Q1) is independently —H, —R¹, —Cl, —OR¹, —SH, —SR¹, —NH₂,        —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —R², —Cl, —OR², —SH, —SR², —NH₂, —NHR²,        —NR² ₂, or —NR^(RC)R^(RD);        or:    -   —R^(Q1) is independently —R¹, —Cl, —OR¹, —SH, —SR¹, —NH₂, —NHR¹,        —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —H, —R², —Cl, —OR², —SH, —SR², —NH₂,        —NHR², —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:    -   —R^(Q1) is independently —H, —R¹, —Cl, —OR¹, —SH, —SR¹, —NH₂,        —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —R², —Cl, —OR², —SH, —SR², —NH₂, —NHR²,        —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:    -   —R^(Q1) is independently —R¹, —Cl, —OR¹, —SH, —SR¹, —NH₂, —NHR¹,        —NR¹ ₂, or —NR^(RA)R^(RB); and    -   —R^(Q2) is independently —H, —R², —Cl, —OR², —SH, —SR², —NH₂,        —NHR², —NR² ₂, or —NR^(RC)R^(RD).        In one embodiment:        either:    -   —R^(Q1) is independently —H, -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino; and    -   —R^(Q2) is independently -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino;        or:    -   —R^(Q1) is independently -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino; and    -   —R^(Q2) is independently —H, -Me, —NH₂, —NHMe, morpholino, or        piperazino, or N-methyl-piperazino.

The Groups —R¹ and —R²

In one embodiment, each —R¹, if present, is independently saturatedaliphatic C₁₋₆alkyl, and is unsubstituted or substituted, for example,with one or more groups selected from —OH, —OR¹¹, —NH₂, —NHR¹¹, and—NR¹¹ ₂, wherein each —R¹¹ is independently saturated aliphaticC₁₋₃alkyl.

In one embodiment, each —R¹¹, if present, is independently -Me or -Et.

In one embodiment, each —R¹, if present, is independently saturatedaliphatic C₁₋₆alkyl, and is unsubstituted.

In one embodiment, each —R¹, if present, is independently saturatedaliphatic C₁₋₄alkyl, and is unsubstituted.

In one embodiment, each —R², if present, is independently saturatedaliphatic C₁₋₆alkyl, and is unsubstituted or substituted, for example,with one or more groups selected from —OH, —OR²², —NH₂, —NHR²², and—NR²² ₂, wherein each —R²² is independently saturated aliphaticC₁₋₃alkyl.

In one embodiment, each —R²², if present, is independently -Me or -Et.

In one embodiment, each —R², if present, is independently saturatedaliphatic C₁₋₆alkyl, and is unsubstituted.

In one embodiment, each —R², if present, is independently saturatedaliphatic C₁₋₄alkyl, and is unsubstituted.

The Groups —R^(1X) and —R^(2X)

In one embodiment, each —R^(1X), if present, is independently saturatedaliphatic C₁₋₄alkyl substituted with one or more groups selected from—F, —Cl, —Br, and —I.

In one embodiment, each —R^(1X), if present, is independently saturatedaliphatic C₁₋₄alkyl substituted with one or more groups selected from —For —Cl.

In one embodiment, each —R^(1X), if present, is independently —CF₃ or—CH₂Br.

In one embodiment, each —R^(1X), if present, is independently —CF₃.

In one embodiment, each —R^(2X), if present, is independently saturatedaliphatic C₁₋₄alkyl substituted with one or more groups selected from—F, —Cl, —Br, and —I.

In one embodiment, each —R^(2X), if present, is independently saturatedaliphatic C₁₋₄alkyl substituted with one or more groups selected from —For —Cl.

In one embodiment, each —R^(2X), if present, is independently —CF₃ or—CH₂Br.

In one embodiment, each —R^(2X), if present, is independently —CF₃.

The Groups —NR^(RA)R^(RB) and —NR^(RC)R^(RD)

In one embodiment:

-   -   —NR^(RA)R^(RB), if present, is independently azetidino,        pyrrolidino, piperidino, piperazino, morpholino, azepino, or        diazepino, and is optionally substituted with one or more groups        selected from saturated aliphatic C₁₋₄alkyl; and    -   —NR^(RC)R^(RD), if present, is independently azetidino,        pyrrolidino, piperidino, piperazino, morpholino, azepino, or        diazepino, and is optionally substituted with one or more groups        selected from saturated aliphatic C₁₋₄alkyl.        In one embodiment:    -   —NR^(RA)R^(RB), if present, is independently piperidino,        piperazino, or morpholino, and is optionally substituted with        one or more groups selected from saturated aliphatic C₁₋₄alkyl;        and    -   —NR^(RC)R^(RD), if present, is independently piperidino,        piperazino, or morpholino, and is optionally substituted with        one or more groups selected from saturated aliphatic C₁₋₄alkyl.

The Group —X—

In one embodiment, —X— is independently —O—, —S—, —S(═O)—, or —S(═O)₂—.

In one embodiment, —X— is independently —O— or —S—.

In one embodiment, —X— is independently —O—.

In one embodiment, —X— is independently —S—.

The Group -M-

In one embodiment, -M- is independently selected from:

In one embodiment, -M- is independently:

In one embodiment, -M- is independently:

In one embodiment, -M- is independently:

In one embodiment, n is independently 0, 1 or 2.

In one embodiment, n is independently 0 or 1.

In one embodiment, n is independently 0.

In one embodiment, n is independently 1.

In one embodiment, -M- is independently:

In one embodiment, -M- is independently:

In one embodiment, -M- is independently:

In one embodiment, -M- is independently:

In one embodiment, each —R^(PH1), if present, is independently —F, —Cl,—Br, —I, —R³, —R^(3Y), —CF₃, —OH, —OR³, —OCF₃, —NH₂, —NHR³, —NR³ ₂, —CN,—SH, or —SR³; wherein each —R³ is independently saturated aliphaticC₁₋₄alkyl, and each —R^(3Y) is independently aliphatic C₂₋₆alkenyl oraliphatic C₂₋₆alkynyl.

In one embodiment, each —R^(PH1), if present, is independently —F, —Cl,—Br, —I, —R³, —OH, —OR³, —SH, or —SR³; wherein each —R³ is independentlysaturated aliphatic C₁₋₄alkyl.

In one embodiment, each —R^(PH1), if present, is independently —F or—SR³.

In one embodiment, each —R^(PH1), if present, is independently —F or—SMe.

In one embodiment, each —R^(PH1), if present, is independently —F.

In one embodiment, each —R^(PH1), if present, is independently —SR³.

In one embodiment, each —R^(PH1), if present, is independently —SMe.

In one embodiment, -M- is independently:

In one embodiment, -M- is independently:

In one embodiment, -M- is independently:

The Group -L-

In one embodiment, J-L- is independently selected from:

-   -   J-NR^(N1)—C(═Y)—NR^(N1)—,    -   J-NR^(N1)—C(═Y)—, and    -   J-C(═Y)—NR^(N1)—.

In one embodiment, ═Y is independently ═O.

In one embodiment, ═Y is independently ═S.

In one embodiment, -L- is independently selected from:

In one embodiment, -L- is independently:

In one embodiment, -L- is independently:

In one embodiment, -L- is independently:

In one embodiment, each —R^(N1), if present, is independently —H orsaturated aliphatic C₁₋₄alkyl.

In one embodiment, each —R^(N1), if present, is independently —H.

The Group -J

In one embodiment, -J is independently phenyl or C₅₋₆heteroaryl, and isoptionally substituted.

In one embodiment, -J is independently phenyl, pyrazolyl, or pyridyl,and is optionally substituted.

In one embodiment, -J is independently phenyl or pyrazolyl, and isoptionally substituted.

In one embodiment, -J is independently phenyl, and is optionallysubstituted.

In one embodiment, -J is independently pyrazolyl, and is optionallysubstituted.

In one embodiment, -J is independently 1H-pyrazol-5-yl, and isoptionally substituted.

In one embodiment, -J is independently pyridyl, and is optionallysubstituted.

In one embodiment, -J is independently pyrid-3-yl, and is optionallysubstituted.

The Group -J: Optional Substituents

In one embodiment, -J is optionally substituted with one or moresubstituents selected from:

-   -   —F, —Cl, —Br, —I, —CF₃, —OCF₃,    -   —R⁴, —R^(4S), —R^(4A), —R^(4B), —R^(4C), -L⁴-R^(4C), —Ar,        -L⁴-Ar,    -   —OH, —OR⁴, -L⁴-OH, -L⁴-OR⁴, —O-L⁴-OH, —O-L⁴-OR⁴,    -   —OR^(4C), —O-L⁴-R^(4C), —OAr, —O-L⁴-Ar,    -   —SH, —SR⁴, —CN, —NO₂,    -   —NH₂, —NHR^(4SS), —R^(N),    -   -L⁴-NH₂, -L⁴-NHR^(4SS), -L⁴-R^(N),    -   —O-L⁴-NH₂, —O-L⁴-NHR^(4SS), —O-L⁴-R^(N),    -   —NH-L⁴-NH₂, —NH-L⁴-NHR^(4SS), —NH-L⁴-RN,    -   —NR⁴-L⁴-NH₂, —NR⁴-L⁴-NHR^(4SS), —NR⁴-L⁴-R^(N),

In one embodiment, -J is optionally substituted with one or moresubstituents selected from:

-   -   —F, —Cl, —Br, —I, —CF₃, —OCF₃,    -   —R⁴, —R^(4S), —R^(4A), —R^(4B), —R^(4C), -L⁴-R^(4C), —Ar,        -L⁴-Ar,    -   —OH, —OR⁴, -L⁴-OH, -L⁴-OR⁴, —O-L⁴-OH, —O-L⁴-OR⁴,    -   —OR^(4C), —O-L⁴-R^(4C), —OAr, —O-L⁴-Ar,    -   —NH₂, —NHR^(4SS), —R^(N),    -   -L⁴-NH₂, -L⁴-NHR^(4SS), -L⁴-R^(N),    -   —O-L⁴-NH₂, —O-L⁴-NHR^(4SS), —O-L⁴-R^(N),    -   —NH-L⁴-NH₂, —NH-L⁴-NHR^(4SS), —NH-L⁴-R^(N),    -   —NR⁴-L⁴-NH₂, —NR⁴-L⁴-NHR^(4SS), and —NR⁴-L⁴-R^(N).

In one embodiment, -J is optionally substituted with one or moresubstituents selected from —F, —Cl, —Br, —I, —R⁴, —Ar, -L⁴-Ar, —OH,—OR⁴, —CF₃, —OCF₃, —OAr, —O-L⁴-Ar.

In one embodiment, each —Ar, if present, is independently optionallysubstituted phenyl or pyridyl, for example, phenyl or pyridyl optionallysubstituted with one or more substituents selected from —F, —Cl, —Br,—I, —R⁵, —OH, —OR⁵, —CF₃, —OCF₃, and —S(═O)₂R⁵.

In one embodiment, -J is optionally substituted with one or moresubstituents selected from —F, —Cl, —Br, —I, —R⁴, —OH, —OR⁴, —CF₃,—OCF₃, and -Ph, wherein each —R⁴ is independently saturated aliphaticC₁₋₄alkyl; and each -Ph denotes optionally substituted phenyl, forexample, phenyl optionally substituted with one or more substituentsselected from —F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵, —CF₃, and —OCF₃, whereineach —R⁵ is independently saturated aliphatic C₁₋₄alkyl.

The Group -J: Substituted Pyrazolyl

In one embodiment, -J is independently pyrazolyl, and is optionallysubstituted.

In one embodiment, -J is independently 1H-pyrazol-5-yl, and isoptionally substituted.

In one embodiment, -J is independently:

wherein:

-   -   —R^(PY1) is independently selected from —R⁴, —R^(4S), —R^(4A),        —R^(4B), R^(4C), -L⁴⁻R^(4C), —Ar, and -L⁴-Ar; and    -   —R^(PY2) is independently —F, —Cl, —Br, —I, —R⁴, —OH, —OR⁴,        —CF₃, —OCF₃, and —Ar.

In one embodiment, —R^(PY1) is independently —Ar.

In one embodiment, —R^(PY1) is independently phenyl or C₅₋₆heteroaryl,and is optionally substituted, for example, with one or moresubstituents selected from —F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵, —CF₃,—OCF₃, and —S(═O)₂R⁵.

In one embodiment, —R^(PY1) is independently phenyl or pyridyl, and isoptionally substituted, for example, with one or more substituentsselected from —F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵, —CF₃, —OCF₃, and—S(═O)₂R⁵.

In one embodiment, -J is independently:

wherein:

-   -   —R^(PY1) is independently phenyl or C₅₋₆heteroaryl, and is        optionally substituted, for example, with one or more        substituents selected from —F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵,        —CF₃, —OCF₃, wherein each —R⁵ is independently saturated        aliphatic C₁₋₄alkyl;    -   —R^(PY2) is independently —F, —Cl, —Br, —I, —R⁴, —OH, —OR⁴,        —CF₃, —OCF₃, and -Ph, wherein each —R⁴ is independently        saturated aliphatic C₁₋₄alkyl.

In one embodiment, —R^(PY1) is independently phenyl or pyridyl, and isoptionally substituted, for example, with one or more substituentsselected from —F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵, —CF₃, —OCF₃.

In one embodiment, —R^(PY1) is independently phenyl, and is optionallysubstituted, for example, with one or more substituents selected from—F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵, —CF₃, —OCF₃.

In one embodiment, —R^(PY1) is independently phenyl, and is optionallysubstituted, for example, with one or more substituents selected from—F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵,

In one embodiment, —R^(PY1) is independently phenyl, and is optionallysubstituted, for example, with one or more substituents selected from—R⁵.

In one embodiment, —R^(PY1) is independently pyridyl, and is optionallysubstituted, for example, with one or more substituents selected from—F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵, —CF₃, —OCF₃.

In one embodiment, —R^(PY1) is independently pyridyl, and is optionallysubstituted, for example, with one or more substituents selected from—OH and —OR⁵.

In one embodiment, each —R⁵, if present, is -Me.

In one embodiment, —R^(PY2) is independently —R⁴.

In one embodiment, —R^(PY2) is independently -tBu.

In one embodiment, -J is independently:

In one embodiment, -J is independently selected from:

In one embodiment, -J is independently selected from:

The Group -J: Phenyl and Substituted Phenyl

In one embodiment, -J is independently phenyl, and is optionallysubstituted.

In one embodiment, -J is independently:

wherein:

-   -   m is independently 0, 1, 2, or 3;    -   each —R^(PH2) is independently selected from:    -   —F, —Cl, —Br, —I, —CF₃, —OCF₃,    -   —R⁴, —R^(4S), —R^(4A), —R^(4B), —R^(4C), -L⁴-R^(4C), —Ar,        -L⁴-Ar,    -   —OH, —OR¹, -L⁴-OH, -L⁴-OR⁴, —O-L⁴-OH, —O-L⁴-OR⁴,    -   —OR^(4C), —O-L⁴-R^(4C), —OAr, —O-L⁴-Ar,    -   —SH, —SR¹, —CN, —NO₂,    -   —NH₂, —NHR^(4SS), —R^(N),    -   -L⁴-NH₂, -L⁴-NHR^(4SS), -L⁴-R^(N),    -   —O-L⁴-NH₂, —O-L⁴-NHR^(4SS), —O-L⁴-R^(N),    -   —NH-L⁴-NH₂, —NH-L⁴-NHR^(4SS), —NH-L⁴-R^(N),    -   —NR⁴-L⁴-NH₂, —NR⁴-L⁴-NHR^(4SS), and —NR⁴-L⁴-R^(N).

In one embodiment, each —R^(PH2), if present, is independently selectedfrom:

-   -   —F, —Cl, —Br, —I, —CF₃, —OCF₃,    -   —R⁴, —R^(4S), —R^(4A), —R^(4B), —R^(4C), -L⁴⁻R^(4C), —Ar,        -L⁴-Ar,    -   —OH, —OR¹, -L⁴-OH, -L⁴-OR⁴, —O-L⁴-OH, —O-L⁴-OR⁴,    -   —OR^(4C), —O-L⁴-R^(4C), —OAr, —O-L⁴-Ar,    -   —NH₂, —NHR^(4SS), —R^(N),    -   -L⁴-NH₂, -L⁴-NHR^(4SS), -L⁴-R^(N),    -   —O-L⁴-NH₂, —O-L⁴-NHR^(4SS), —O-L⁴-R^(N),    -   —NH-L⁴-NH₂, —NH-L⁴-NHR^(4SS), —NH-L⁴-R^(N),    -   —NR⁴-L⁴-NH₂, —NR⁴-L⁴-NHR^(4SS), and —NR⁴-L⁴-R^(N).

In one embodiment, each —R^(PH2), if present, is independently selectedfrom:

-   -   —F, —Cl, —Br, —I, —CF₃, —OCF₃,    -   —R⁴, —R^(4S), —Ar, -L⁴-Ar,    -   —OH, —OR⁴, —OAr, —O-L⁴-Ar, -L⁴-OH, -L⁴-OR⁴, —O-L⁴-OH, —O-L⁴-OR⁴,    -   —NH₂, —NHR^(4SS), —R^(N),    -   -L⁴-NH₂, -L⁴-NHR^(4SS), -L⁴-R^(N),    -   —O-L⁴-NH₂, —O-L⁴-NHR^(4SS), —O-L⁴-R^(N),    -   —NH-L⁴-NH₂, —NH-L⁴-NHR^(4SS), —NH-L⁴-R^(N),    -   —NR⁴-L⁴-NH₂, —NR⁴-L⁴-NHR^(4SS), and —NR⁴-L⁴-R^(N).

In one embodiment, each —R^(PH2), if present, is independently selectedfrom:

-   -   —F, —Cl, —Br, —I, —CF₃, —OCF₃,    -   —R⁴, —R^(4S), —Ar, -L⁴-Ar,    -   —OH, —OR⁴, —OAr, —O-L⁴-Ar,    -   —NH₂, —NHR^(4SS), and —R^(N).

In one embodiment, each —R^(PH2), if present, is independently selectedfrom:

-   -   —F, —Cl, —Br, —I, —CF₃, —OCF₃,    -   R⁴, —R^(4S),    -   —OH, —OR⁴,    -   —NH₂, —NHR^(4SS), and —R^(N).

In one embodiment, -J is independently:

wherein:

-   -   m is independently 0, 1, 2, or 3;    -   each —R^(PH2) is independently —F, —Cl, —Br, —I, —R⁴, —OH, —OR⁴,        —CF₃, or —OCF₃, wherein each —R⁴ is independently saturated        aliphatic C₁₋₄alkyl.

In one embodiment, m is independently 0, 1, or 2.

In one embodiment, m is independently 1 or 2.

In one embodiment, m is independently 1.

In one embodiment, m is independently 2.

In one embodiment, each —R^(PH2), if present, is independently —F, —Cl,-tBu, —CF₃, or —OCF₃.

Combinations

Each and every compatible combination of the embodiments described aboveis explicitly disclosed herein, as if each and every combination wasindividually and explicitly recited.

Examples of Specific Embodiments

In one embodiment, the compounds are selected from compounds of thefollowing formulae and pharmaceutically acceptable salts, hydrates, andsolvates thereof:

Cmpd. Structure AA-001

AA-002

AA-003

AA-004

AA-005

AA-006

AA-007

AA-008

AA-009

AA-010

AA-011

AA-012

AA-013

AA-014

AA-015

AA-016

AA-017

AA-018

AA-019

AA-020

AA-021

AA-022

AA-023

AA-024

AA-025

AA-026

AA-027

AA-028

AA-029

AA-030

AA-031

AA-032

AA-033

AA-034

AA-035

AA-036

AA-037

AA-038

AA-039

AA-040

AA-041

AA-042

AA-043

AA-044

AA-045

AA-046

AA-047

AA-048

AA-049

AA-050

AA-051

AA-052

AA-053

AA-054

AA-055

AA-056

AA-057

AA-058

AA-059

AA-060

AA-061

AA-062

AA-063

AA-064

AA-065

AA-066

AA-067

AA-068

AA-069

AA-070

AA-071

AA-072

AA-073

AA-074

AA-075

AA-076

AA-077

AA-078

AA-079

AA-080

AA-081

AA-082

AA-083

AA-084

AA-085

AA-086

AA-087

AA-088

AA-089

AA-090

AA-091

AA-092

AA-093

AA-094

AA-095

AA-096

AA-097

AA-098

AA-099

AA-100

Substantially Purified Forms

One aspect of the present invention pertains to PDP8 compounds, asdescribed herein, in substantially purified form and/or in a formsubstantially free from contaminants.

In one embodiment, the substantially purified form is at least 50% byweight, e.g., at least 60% by weight, e.g., at least 70% by weight,e.g., at least 80% by weight, e.g., at least 90% by weight, e.g., atleast 95% by weight, e.g., at least 97% by weight, e.g., at least 98% byweight, e.g., at least 99% by weight.

Unless specified, the substantially purified form refers to the compoundin any stereoisomeric or enantiomeric form. For example, in oneembodiment, the substantially purified form refers to a mixture ofstereoisomers, i.e., purified with respect to other compounds. In oneembodiment, the substantially purified form refers to one stereoisomer,e.g., optically pure stereoisomer. In one embodiment, the substantiallypurified form refers to a mixture of enantiomers. In one embodiment, thesubstantially purified form refers to a equimolar mixture of enantiomers(i.e., a racemic mixture, a racemate). In one embodiment, thesubstantially purified form refers to one enantiomer, e.g., opticallypure enantiomer.

In one embodiment, the contaminants represent no more than 50% byweight, e.g., no more than 40% by weight, e.g., no more than 30% byweight, e.g., no more than 20% by weight, e.g., no more than 10% byweight, e.g., no more than 5% by weight, e.g., no more than 3% byweight, e.g., no more than 2% by weight, e.g., no more than 1% byweight.

Unless specified, the contaminants refer to other compounds, that is,other than stereoisomers or enantiomers. In one embodiment, thecontaminants refer to other compounds and other stereoisomers. In oneembodiment, the contaminants refer to other compounds and the otherenantiomer.

In one embodiment, the substantially purified form is at least 60%optically pure (i.e., 60% of the compound, on a molar basis, is thedesired stereoisomer or enantiomer, and 40% is the undesiredstereoisomer or enantiomer), e.g., at least 70% optically pure, e.g., atleast 80% optically pure, e.g., at least 90% optically pure, e.g., atleast 95% optically pure, e.g., at least 97% optically pure, e.g., atleast 98% optically pure, e.g., at least 99% optically pure.

Isomers

Certain compounds may exist in one or more particular geometric,optical, enantiomeric, diasteriomeric, epimeric, atropic,stereoisomeric, tautomeric, conformational, or anomeric forms, includingbut not limited to, cis- and trans-forms; E- and Z-forms; c-, t-, andr-forms; endo- and exo-forms; R-, S-, and meso-forms; D- and L-forms; d-and l-forms; (+) and (−) forms; keto-, enol-, and enolate-forms; syn-and anti-forms; synclinal- and anticlinal-forms; α- and β-forms; axialand equatorial forms; boat-, chair-, twist-, envelope-, andhalfchair-forms; and combinations thereof, hereinafter collectivelyreferred to as “isomers” (or “isomeric forms”).

Note that, except as discussed below for tautomeric forms, specificallyexcluded from the term “isomers,” as used herein, are structural (orconstitutional) isomers (i.e., isomers which differ in the connectionsbetween atoms rather than merely by the position of atoms in space). Forexample, a reference to a methoxy group, —OCH₃, is not to be construedas a reference to its structural isomer, a hydroxymethyl group, —CH₂OH.Similarly, a reference to ortho-chlorophenyl is not to be construed as areference to its structural isomer, meta-chlorophenyl. However, areference to a class of structures may well include structurallyisomeric forms falling within that class (e.g., C₁₋₇alkyl includesn-propyl and iso-propyl; butyl includes n-, iso-, sec-, and tert-butyl;methoxyphenyl includes ortho-, meta-, and para-methoxyphenyl).

The above exclusion does not pertain to tautomeric forms, for example,keto-, enol-, and enolate-forms, as in, for example, the followingtautomeric pairs: keto/enol (illustrated below), imine/enamine,amide/imino alcohol, amidine/amidine, nitroso/oxime,thioketone/enethiol, N-nitroso/hydroxyazo, and nitro/aci-nitro.

Note that specifically included in the term “isomer” are compounds withone or more isotopic substitutions. For example, H may be in anyisotopic form, including ¹H, ²H (D), and ³H (T); C may be in anyisotopic form, including ¹²C, ¹³C, and ¹⁴C; O may be in any isotopicform, including ¹⁶O and ¹⁸O; and the like.

Unless otherwise specified, a reference to a particular compoundincludes all such isomeric forms, including mixtures (e.g., racemicmixtures) thereof. Methods for the preparation (e.g., asymmetricsynthesis) and separation (e.g., fractional crystallisation andchromatographic means) of such isomeric forms are either known in theart or are readily obtained by adapting the methods taught herein, orknown methods, in a known manner.

Salts

It may be convenient or desirable to prepare, purify, and/or handle acorresponding salt of the compound, for example, apharmaceutically-acceptable salt. Examples of pharmaceuticallyacceptable salts are discussed in Berge et al., 1977, “PharmaceuticallyAcceptable Salts,” J. Pharm. Sci., Vol. 66, pp. 1-19.

For example, if the compound is anionic, or has a functional group whichmay be anionic (e.g., —COOH may be —COO⁻), then a salt may be formedwith a suitable cation. Examples of suitable inorganic cations include,but are not limited to, alkali metal ions such as Na⁺ and K⁺, alkalineearth cations such as Ca²⁺ and Mg²⁺, and other cations such as Al⁺³.Examples of suitable organic cations include, but are not limited to,ammonium ion (i.e., NH₄ ⁺) and substituted ammonium ions (e.g., NH₃R⁺,NH₂R₂ ⁺, NHR₃ ⁺, NR₄ ⁺). Examples of some suitable substituted ammoniumions are those derived from: ethylamine, diethylamine,dicyclohexylamine, triethylamine, butylamine, ethylenediamine,ethanolamine, diethanolamine, piperazine, benzylamine,phenylbenzylamine, choline, meglumine, and tromethamine, as well asamino acids, such as lysine and arginine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

If the compound is cationic, or has a functional group which may becationic (e.g., —NH₂ may be —NH₃ ⁺), then a salt may be formed with asuitable anion. Examples of suitable inorganic anions include, but arenot limited to, those derived from the following inorganic acids:hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric,nitrous, phosphoric, and phosphorous.

Examples of suitable organic anions include, but are not limited to,those derived from the following organic acids: 2-acetyoxybenzoic,acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric,edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucheptonic,gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalenecarboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic,methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic,phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic,succinic, sulfanilic, tartaric, toluenesulfonic, and valeric. Examplesof suitable polymeric organic anions include, but are not limited to,those derived from the following polymeric acids: tannic acid,carboxymethyl cellulose.

Unless otherwise specified, a reference to a particular compound alsoincludes salt forms thereof.

Solvates and Hydrates

It may be convenient or desirable to prepare, purify, and/or handle acorresponding solvate of the compound. The term “solvate” is used hereinin the conventional sense to refer to a complex of solute (e.g.,compound, salt of compound) and solvent. If the solvent is water, thesolvate may be conveniently referred to as a hydrate, for example, amono-hydrate, a di-hydrate, a tri-hydrate, etc.

Unless otherwise specified, a reference to a particular compound alsoincludes solvate and hydrate forms thereof.

Chemically Protected Forms

It may be convenient or desirable to prepare, purify, and/or handle thecompound in a chemically protected form. The term “chemically protectedform” is used herein in the conventional chemical sense and pertains toa compound in which one or more reactive functional groups are protectedfrom undesirable chemical reactions under specified conditions (e.g.,pH, temperature, radiation, solvent, and the like). In practice, wellknown chemical methods are employed to reversibly render unreactive afunctional group, which otherwise would be reactive, under specifiedconditions. In a chemically protected form, one or more reactivefunctional groups are in the form of a protected or protecting group(also known as a masked or masking group or a blocked or blockinggroup). By protecting a reactive functional group, reactions involvingother unprotected reactive functional groups can be performed, withoutaffecting the protected group; the protecting group may be removed,usually in a subsequent step, without substantially affecting theremainder of the molecule. See, for example, Protective Groups inOrganic Synthesis (T. Green and P. Wuts; 3rd Edition; John Wiley andSons, 1999).

A wide variety of such “protecting,” “blocking,” or “masking” methodsare widely used and well known in organic synthesis. For example, acompound which has two nonequivalent reactive functional groups, both ofwhich would be reactive under specified conditions, may be derivatizedto render one of the functional groups “protected,” and thereforeunreactive, under the specified conditions; so protected, the compoundmay be used as a reactant which has effectively only one reactivefunctional group. After the desired reaction (involving the otherfunctional group) is complete, the protected group may be “deprotected”to return it to its original functionality.

For example, a hydroxy group may be protected as an ether (—OR) or anester (—OC(═O)R), for example, as: a t-butyl ether; a benzyl, benzhydryl(diphenylmethyl), or trityl (triphenylmethyl)ether; a trimethylsilyl ort-butyldimethylsilyl ether; or an acetyl ester (—OC(═O)CH₃, —OAc).

For example, an aldehyde or ketone group may be protected as an acetal(R—CH(OR)₂) or ketal (R₂C(OR)₂), respectively, in which the carbonylgroup (>C═O) is converted to a diether (>C(OR)₂), by reaction with, forexample, a primary alcohol. The aldehyde or ketone group is readilyregenerated by hydrolysis using a large excess of water in the presenceof acid.

For example, an amine group may be protected, for example, as an amide(—NRCO—R) or a urethane (—NRCO—OR), for example, as: a methyl amide(—NHCO—CH₃); a benzyloxy amide (—NHCO—OCH₂C₆H₅, —NH-Cbz); as a t-butoxyamide (—NHCO—OC(CH₃)₃, —NH-Boc); a 2-biphenyl-2-propoxy amide(—NHCO—OC(CH₃)₂C₆H₄C₆H₅, —NH-Bpoc), as a 9-fluorenylmethoxy amide(—NH-Fmoc), as a 6-nitroveratryloxy amide (—NH-Nvoc), as a2-trimethylsilylethyloxy amide (—NH-Teoc), as a 2,2,2-trichloroethyloxyamide (—NH-Troc), as an allyloxy amide (—NH-Alloc), as a2(-phenylsulfonyl)ethyloxy amide (—NH-Psec); or, in suitable cases(e.g., cyclic amines), as a nitroxide radical (>N—O.).

For example, a carboxylic acid group may be protected as an ester forexample, as: an C₁₋₇alkyl ester (e.g., a methyl ester; a t-butyl ester);a C₁₋₇haloalkyl ester (e.g., a C₁₋₇-trihaloalkyl ester); atriC₁₋₇alkylsilyl-C₁₋₇alkyl ester; or a C₅₋₂₀aryl-C₁₋₇alkyl ester (e.g.,a benzyl ester; a nitrobenzyl ester); or as an amide, for example, as amethyl amide.

For example, a thiol group may be protected as a thioether (—SR), forexample, as: a benzyl thioether; an acetamidomethyl ether(—S—CH₂NHC(═O)CH₃).

Prodrugs

It may be convenient or desirable to prepare, purify, and/or handle thecompound in the form of a prodrug. The term “prodrug,” as used herein,pertains to a compound which, when metabolised (e.g., in vivo), yieldsthe desired active compound. Typically, the prodrug is inactive, or lessactive than the desired active compound, but may provide advantageoushandling, administration, or metabolic properties.

For example, some prodrugs are esters of the active compound (e.g., aphysiologically acceptable metabolically labile ester). Duringmetabolism, the ester group (—C(═O)OR) is cleaved to yield the activedrug. Such esters may be formed by esterification, for example, of anyof the carboxylic acid groups (—C(═O)OH) in the parent compound, with,where appropriate, prior protection of any other reactive groups presentin the parent compound, followed by deprotection if required.

Also, some prodrugs are activated enzymatically to yield the activecompound, or a compound which, upon further chemical reaction, yieldsthe active compound (for example, as in ADEPT, GDEPT, LIDEPT, etc.). Forexample, the prodrug may be a sugar derivative or other glycosideconjugate, or may be an amino acid ester derivative.

Chemical Synthesis

Several methods for the chemical synthesis of PDP8 compounds of thepresent invention are described herein. These and/or other well knownmethods may be modified and/or adapted in known ways in order tofacilitate the synthesis of additional compounds within the scope of thepresent invention.

Compositions

One aspect of the present invention pertains to a composition (e.g., apharmaceutical composition) comprising a PDP8 compound, as describedherein, and a pharmaceutically acceptable carrier, diluent, orexcipient.

Another aspect of the present invention pertains to a method ofpreparing a composition (e.g., a pharmaceutical composition) comprisingadmixing a PDP8 compound, as described herein, and a pharmaceuticallyacceptable carrier, diluent, or excipient.

Uses

The compounds described herein are useful, for example, in the treatmentof diseases and disorders that are ameliorated by the inhibition of RAF(e.g., B-RAF), such as, for example, proliferative disorders, cancer,etc.

Use in Methods of Inhibiting RAF (e.g., B-RAF)

One aspect of the present invention pertains to a method of inhibitingRAF (e.g., B-RAF) function, in vitro or in vivo, comprising contacting aRAF (e.g., B-RAF) with an effective amount of a PDP8 compound, asdescribed herein.

One aspect of the present invention pertains to a method of inhibitingRAF (e.g., B-RAF) function in a cell, in vitro or in vivo, comprisingcontacting the cell with an effective amount of a PDP8 compound, asdescribed herein.

In one embodiment, the method is performed in vitro.

In one embodiment, the method is performed in vivo.

One of ordinary skill in the art is readily able to determine whether ornot, and/or the degree to which, a candidate compound inhibits RAF(e.g., B-RAF) function. Suitable assays for determining RAF (e.g.,B-RAF) function inhibition are described herein and/or are known in theart.

B-RAF Assays:

B-raf kinase activity is measured using a 4-tiered cascade enzyme assaysimilar to that described by Marais R., et al., 1997, J. Biol. Chem.,Vol. 272, pp. 4378-4383. B-Raf containing the V600E mutation (Davies,H., et al., 2002, Nature, Vol. 417, pp. 949-954) and an N-terminalMDRGSH6 tag is expressed in SF9 insect cells. Detergent soluble extractsfrom these cells are diluted 1:100 into an assay mixture containingGST-MEK-H6 (6.5 μg/ml) and GST-ERK-H6 (100 μg/ml) in a buffer containing800 μM ATP and appropriate concentrations of inhibitor or diluent ascontrol. The mixture is incubated for up to 10 minutes at 30° C. toactivate the ERK in a B-Raf dependent manner within the cascade. Thereaction is then stopped by addition of 20 mM EDTA. The extent ofactivation of the GST-ERK is then determined by adding a portion of thisquenched reaction mixture to a further reaction mixture containing MBPand 100 μM ATP/gamma [³²P]ATP. After 12 minutes' incubation at 30° C.,the incorporation of [³²P] into the MBP substrate, as a measure of B-rafactivity, is determined by precipitation with phosphoric acid andisolation by filtration on p81 phosphocellulose paper. The % inhibitionof the B-raf kinase activity is calculated and plotted in order todetermine the concentration of test compound required to inhibit 50% ofthe B-raf kinase activity (IC₅₀).

Alternatively, B-raf kinase activity is measured using a different4-tiered cascade enzyme assay. B-Raf containing the V600E mutation(Davies, H., et al., 2002, Nature, Vol. 417, pp. 949-954) and anN-terminal MDRGSH6 tag is expressed in SF9 insect cells. Detergentsoluble extracts from these cells are diluted 1:250 into an assaymixture containing GST-MEK-H6 (25 μg/ml), GST-ERK-H6 (281.25 μg/ml) andMBP in a buffer containing appropriate concentrations of inhibitor ordiluent as control. 0.03 μL (100 μM) ATP is added and the mixture isincubated for up to 10 minutes at 30° C. to activate the ERK in a B-Rafdependent manner within the cascade. The extent of activation of theGST-ERK is then determined by adding 0.033 μL (100 μM) HOT ³²Pα. After10 minutes' incubation at 30° C., the reaction is stopped by isolationof a portion of the reaction mixture on p81 phosphocellulose paper andsubmersion of this paper in 0.4% orthophosphoric acid. Incorporation of[³²P] into the MBP substrate, as a measure of B-raf activity, isdetermined using a Packard Cernekov counter. The % inhibition of theB-raf kinase activity is calculated and plotted in order to determinethe concentration of test compound required to inhibit 50% of the B-rafkinase activity (IC₅₀).

C-RAF Assay:

C-raf (human) is diluted to a 10× working stock in 50 mM Tris pH 7.5,0.1 mM EGTA, 0.1 mM sodium vanadate, 0.1% β-mercaptoethanol, 1 mg/mlBSA. One unit equals the incorporation of 1 nmol of phosphate per minuteinto myelin basic protein per minute. In a final reaction volume of 25μl, c-raf (5-10 mU) is incubated with 25 mM Tris pH 7.5, 0.02 mM EGTA,0.66 mg/ml myelin basic protein, 10 mM MgAcetate, [γ-³³P-ATP] (specificactivity approx 500 cpm/pmol, concentration as required) and appropriateconcentrations of inhibitor or diluent as control. The reaction isinitiated by the addition of Mg²+[γ-³³P-ATP]. After incubation for 40minutes at room temperature, the reaction is stopped by the addition of5 μl of a 3% phosphoric acid solution. 10 μl of the reaction is spottedonto a P30 filtermat and washed 3 times for 5 minutes in 75 mMphosphoric acid and once in methanol prior to drying and counting todetermine the C-raf activity. The % inhibition of the C-raf kinaseactivity is calculated and plotted in order to determine theconcentration of test compound required to inhibit 50% of the C-rafkinase activity (IC₅₀).

Selectivity:

In one embodiment, the PDP8 compound selectively inhibits one RAF (e.g.,B-RAF), over at least one other RAF (e.g., A-RAF and/or C-RAF).

For example, in one embodiment, the ratio of the IC₅₀ value for B-RAF tothe IC₅₀ value for the other RAF (e.g., A-RAF and/or C-RAF) is at least10, more preferably at least 100, most preferably at least 1000.

Use in Methods of Inhibiting Cell Proliferation, Etc.

The PDP8 compounds described herein, e.g., (a) regulate (e.g., inhibit)cell proliferation; (b) inhibit cell cycle progression; (c) promoteapoptosis; or (d) a combination of one or more of these.

One aspect of the present invention pertains to a method of regulating(e.g., inhibiting) cell proliferation (e.g., proliferation of a cell),inhibiting cell cycle progression, promoting apoptosis, or a combinationof one or more these, in vitro or in vivo, comprising contacting a cellwith an effective amount of a PDP8 compound, as described herein.

In one embodiment, the method is a method of regulating (e.g.,inhibiting) cell proliferation (e.g., proliferation of a cell), in vitroor in vivo, comprising contacting a cell with an effective amount of aPDP8 compound, as described herein.

In one embodiment, the method is performed in vitro.

In one embodiment, the method is performed in vivo.

In one embodiment, the PDP8 compound is provided in the form of apharmaceutically acceptable composition.

Any type of cell may be treated, including but not limited to, lung,gastrointestinal (including, e.g., bowel, colon), breast (mammary),ovarian, prostate, liver (hepatic), kidney (renal), bladder, pancreas,brain, and skin.

One of ordinary skill in the art is readily able to determine whether ornot a candidate compound regulates (e.g., inhibits) cell proliferation,etc. For example, assays which may conveniently be used to assess theactivity offered by a particular compound are described herein.

For example, a sample of cells (e.g., from a tumour) may be grown invitro and a compound brought into contact with said cells, and theeffect of the compound on those cells observed. As an example of“effect,” the morphological status of the cells (e.g., alive or dead,etc.) may be determined. Where the compound is found to exert aninfluence on the cells, this may be used as a prognostic or diagnosticmarker of the efficacy of the compound in methods of treating a patientcarrying cells of the same cellular type.

Use in Methods of Therapy

Another aspect of the present invention pertains to a PDP8 compound, asdescribed herein, for use in a method of treatment of the human oranimal body by therapy.

Use in the Manufacture of Medicaments

Another aspect of the present invention pertains to use of a PDP8compound, as described herein, in the manufacture of a medicament foruse in treatment.

In one embodiment, the medicament comprises the PDP8 compound.

Methods of Treatment

Another aspect of the present invention pertains to a method oftreatment comprising administering to a patient in need of treatment atherapeutically effective amount of a PDP8 compound, as describedherein, preferably in the form of a pharmaceutical composition.

Conditions Treated—Conditions Ameliorated by the Inhibition of RAF

In one embodiment (e.g., of use in methods of therapy, of use in themanufacture of medicaments, of methods of treatment), the treatment istreatment of a disease or disorder that is characterised by theup-regulation and/or activation of RAF (e.g., B-RAF), and/or isameliorated by the inhibition of RAF (e.g., B-RAF).

In one embodiment, the treatment is treatment of cancer that ischaracterised by the up-regulation and/or activation of RAF (e.g.,B-RAF), and/or is ameliorated by the inhibition of RAF (e.g., B-RAF).

Conditions Treated—Conditions Ameliorated by the Inhibition of RTKs

In one embodiment (e.g., of use in methods of therapy, of use in themanufacture of medicaments, of methods of treatment), the treatment istreatment of a disease or disorder that is characterised by theup-regulation and/or activation of a receptor tyrosine kinase (RTK),and/or is ameliorated by the inhibition of a receptor tyrosine kinase(RTK). Examples of RTKs include FGFR, Tie, VEGFR and/or Eph, forexample, FGFR-1, FGFR-2, FGFR-3, Tie2, VEGFR-2 and/or EphB2.

In one embodiment, the treatment is treatment of cancer that ischaracterised by the up-regulation and/or activation of a receptortyrosine kinase (RTK), and/or is ameliorated by the inhibition of areceptor tyrosine kinase (RTK).

Conditions Treated—Conditions characterised by Angiogenesis

In one embodiment (e.g., of use in methods of therapy, of use in themanufacture of medicaments, of methods of treatment), the treatment istreatment of a disease or disorder that is characterised byinappropriate, excessive, and/or undesirable angiogenesis (as“anti-angiogenesis agents”). Examples of such disorders are discussedherein.

Conditions Treated—Proliferative Disorders and Cancer

The PDP8 compounds are useful in the treatment of proliferativedisorders (as “anti-proliferative agents”), cancer (as “anti-canceragents”), etc.

In one embodiment (e.g., of use in methods of therapy, of use in themanufacture of medicaments, of methods of treatment), the treatment istreatment of a proliferative disorder.

The term “proliferative disorder,” as used herein, pertains to anunwanted or uncontrolled cellular proliferation of excessive or abnormalcells which is undesired, such as, neoplastic or hyperplastic growth.

In one embodiment, the treatment is treatment of: a proliferativedisorder characterised by benign, pre-malignant, or malignant cellularproliferation, including but not limited to, neoplasms, hyperplasias,and tumours (e.g., histocytoma, glioma, astrocyoma, osteoma), cancers(see below), psoriasis, bone diseases, fibroproliferative disorders(e.g., of connective tissues), pulmonary fibrosis, atherosclerosis,smooth muscle cell proliferation in the blood vessels, such as stenosisor restenosis following angioplasty.

In one embodiment, the treatment is treatment of: cancer.

In one embodiment, the treatment is treatment of: lung cancer, smallcell lung cancer, non-small cell lung cancer, gastrointestinal cancer,stomach cancer, bowel cancer, colon cancer, rectal cancer, colorectalcancer, thyroid cancer, breast cancer, ovarian cancer, endometrialcancer, prostate cancer, testicular cancer, liver cancer, kidney cancer,renal cell carcinoma, bladder cancer, pancreatic cancer, brain cancer,glioma, sarcoma, osteosarcoma, bone cancer, skin cancer, squamouscancer, Kaposi's sarcoma, melanoma, malignant melanoma, lymphoma, orleukemia.

In one embodiment, the treatment is treatment of:

-   -   a carcinoma, for example a carcinoma of the bladder, breast,        colon (e.g., colorectal carcinomas such as colon adenocarcinoma        and colon adenoma), kidney, epidermal, liver, lung (e.g.,        adenocarcinoma, small cell lung cancer and non-small cell lung        carcinomas), oesophagus, gall bladder, ovary, pancreas (e.g.,        exocrine pancreatic carcinoma), stomach, cervix, thyroid,        prostate, skin (e.g., squamous cell carcinoma);    -   a hematopoietic tumour of lymphoid lineage, for example        leukemia, acute lymphocytic leukemia, B-cell lymphoma, T-cell        lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell        lymphoma, or Burkett's lymphoma;    -   a hematopoietic tumor of myeloid lineage, for example acute and        chronic myelogenous leukemias, myelodysplastic syndrome, or        promyelocytic leukemia;    -   a tumour of mesenchymal origin, for example fibrosarcoma or        habdomyosarcoma;    -   a tumor of the central or peripheral nervous system, for example        astrocytoma, neuroblastoma, glioma or schwannoma;    -   melanoma; seminoma; teratocarcinoma; osteosarcoma; xenoderoma        pigmentoum; keratoctanthoma; thyroid follicular cancer; or        Kaposi's sarcoma.

In one embodiment, the treatment is treatment of solid tumour cancer.

In one embodiment, the treatment is treatment of melanoma or malignantmelanoma.

In one embodiment, the treatment is treatment of colorectal cancer.

The anti-cancer effect may arise through one or more mechanisms,including but not limited to, the regulation of cell proliferation, theinhibition of cell cycle progression, the inhibition of angiogenesis(the formation of new blood vessels), the inhibition of metastasis (thespread of a tumour from its origin), the inhibition of invasion (thespread of tumour cells into neighbouring normal structures), or thepromotion of apoptosis (programmed cell death). The PDP8 compounds ofthe present invention may be used in the treatment of the cancersdescribed herein, independent of the mechanisms discussed herein.

Conditions Treated—Proliferative Disorders and Cancer Associated withRAF

Cancers with, for example, activating mutations of ras, raf and EGFR orover expression of ras, raf and EGFR including any of the isoformsthereof, may be particularly sensitive to inhibitors of RAF (e.g.,B-RAF) activity. Patients with activating mutants of RAF (e.g., B-RAF)may also find treatment with inhibitors of RAF (e.g., B-RAF) activityparticularly beneficial. Cancers with other abnormalities leading to anupregulated raf-MEK-ERK pathway signal may also be particularlysensitive to treatment with inhibitors of RAF (e.g., B-RAF) activity.Examples of such abnormalities include consitutive activation of agrowth factor receptor; overexpression of one or more growth factorreceptors; and overexpression of one or more growth factors.

In one embodiment (e.g., of use in methods of therapy, of use in themanufacture of medicaments, of methods of treatment), the treatment istreatment of a proliferative disorder as described above, for example,cancer, that is characterised by:

(a) activating mutants of ras or raf;(b) upregulation of ras or raf;(c) upregulated raf-MEK-ERK pathway signals;(d) upregulation of growth factor receptors, such as ERBB2 and EGFR.

In one embodiment, the proliferative disorder is characterised by cellswhich overexpress RAF (e.g., B-RAF) or express or overexpress mutant raf(e.g., B-RAF). In one embodiment, the proliferative disorder ischaracterised by cells which overexpress raf (e.g., B-RAF). In oneembodiment, the proliferative disorder is characterised by cells whichexpress or overexpress mutant RAF (e.g., B-RAF). In one embodiment, theproliferative disorder is characterised by cells which overexpress RAF(e.g., B-RAF), or overexpress mutant RAF (e.g., B-RAF), as compared tocorresponding normal cells. In one embodiment, the overexpression is bya factor of 1.5, 2, 3, 5, 10, or 20.

In one embodiment (e.g., of use in methods of therapy, of use in themanufacture of medicaments, of methods of treatment), the treatment istreatment of a disease or disorder associated with a mutated form of RAF(e.g., B-RAF), such as, for example, the mutations described in Wan, P.,et al., 2004, Cell, Vol. 116, pp. 855-867 and Stratton et al., 2003,published international patent application publication number WO03/056036.

Conditions Treated—Inflammation Etc.

The PDP8 compounds are useful in the treatment of disorders associatedwith inflammation (as “anti-inflammation agents”), etc.

The function of inflammatory cells is controlled by many factors theeffects of which are mediated by different signal transduction pathways.Although some key pro-inflammatory functions are mediated by p38 Mapkinase (e.g., TNF release), others are mediated by other pathways. Theraf-MEK-ERK pathway, in particular, is an important activating andproliferative signal in many inflammatory cells. B and T lymphoctyes, inparticular, require activation of the raf-MEK-ERK pathway for clonalexpansion and generation of effector populations (see, e.g., Cantrell,D. A., 2003, Immunol Rev., Vol. 192, pp. 122-130; Genot, E. andCantrell, D. A., 2000, Curr. Opin. Immunol., Vol. 12(3), pp. 289-294).

In one embodiment, the treatment is treatment of: inflammatory diseases,such as rheumatoid arthritis, osteoarthritis, rheumatoid spondylitis,gouty arthritis, traumatic arthritis, rubella arthritis, psoriaticarthritis, and other arthritic conditions; Alzheimer's disease; toxicshock syndrome, the inflammatory reaction induced by endotoxin orinflammatory bowel disease; tuberculosis; atherosclerosis; muscledegeneration; Reiter's syndrome; gout; acute synovitis; sepsis; septicshock; endotoxic shock; gram negative sepsis; adult respiratory distresssyndrome; cerebral malaria; chronic pulmonary inflammatory disease;silicosis; pulmonary sarcoisosis; bone resorption diseases; reperfusioninjury; graft versus host reaction; allograft rejections; fever andmyalgias due to infection, such as influenza, cachexia, in particularcachexia secondary to infection or malignancy, cachexia secondary toacquired immune deficiency syndrome (AIDS); AIDS; ARC (AIDS relatedcomplex); keloid formation; scar tissue formation; Crohn's disease;ulcerative colitis; pyresis; chronic obstructive pulmonary disease(COPD); acute respiratory distress syndrome (ARDS); asthma; pulmonaryfibrosis; bacterial pneumonia.

In one preferred embodiment, the treatment is treatment of: arthriticconditions, including rheumatoid arthritis and rheumatoid spondylitis;inflammatory bowel disease, including Crohn's disease and ulcerativecolitis; and chronic obstructive pulmonary disease (COPD).

In one preferred embodiment, the treatment is treatment of: aninflammatory disorder characterized by T-cell proliferation (T-cellactivation and growth), for example, tissue graft rejection, endotoxinshock, and glomerular nephritis.

Screening

Prior to treatment, a patient may be screened to determine whether adisease or disorder from which the patient is or may be suffering is onewhich would be susceptible to treatment with a compound that inhibitsRAF (e.g., B-RAF) activity or has activity against an RTK (e.g., FGFR-1,FGFR-2, FGFR-3, VEGFR-2, Tie2, EphB2).

For example, a biological sample taken from a patient may be analysed todetermine whether a disease or disorder, such as cancer, that thepatient is or may be suffering from is one which is characterised byelevated expression or activation of RAF (e.g., B-RAF), or an RTK (e.g.,FGFR-1, FGFR-2, FGFR-3, VEGFR-2, Tie2, EphB2), or is the result of anactivating mutation. Thus, the patient may be subjected to a diagnostictest to detect a marker characteristic of over-expression or activationof RAF (e.g., B-RAF) or an RTK (e.g., FGFR-1, FGFR-2, FGFR-3, VEGFR-2,Tie2, EphB2), or a mutation thereof.

As used herein, the term “marker” includes genetic markers (including,e.g., the measurement of DNA composition to identify mutations of raf,ras, MEK, ERK or a growth factor such as ERBB2 or EGFR) and markerswhich are characteristic of upregulation of raf, ras, MEK, ERK, growthfactors receptors such as ERBB2 or EGFR including enzyme activity,enzyme levels, enzyme state (e.g. phosphorylated or not) and mRNA levelsof the aforementioned proteins. Methods for identification and analysisof mutations are well known. See, for example, Anticancer Research,1999, Vol. 19(4A), pp. 2481-2483; Clin. Chem., 2002, Vol. 48, p. 428;Cancer Research, 2003, Vol. 63(14), pp. 3955-3957.

The term “marker” further includes genetic markers including, forexample, the measurement of DNA composition to identify mutations ofRTKs, e.g., FGFR-1, FGFR-2, FGFR-3, VEGFR-2, Tie2, and EphB2. The term“marker” also includes markers that are characteristic of up-regulationof RTKs, including enzyme activity, enzyme levels, enzyme state (e.g.,phosphorylated or not) and mRNA levels of the aforementioned proteins.

Upregulation includes elevated expression or over expression, includinggene amplification (i.e., multiple gene copies), increased expression bya transcriptional effect, hyperactivity, and activation, includingactivation by mutations.

Other tumours that have an upregulated raf-MEK-ERK pathway signal mayalso be particularly sensitive to inhibitors of RAF (e.g., B-RAF)activity. A number of assays exist which can identify tumours thatexhibit upregulation in the raf-MEK-ERK pathway, including thecommercially available MEK1/2 (MAPK Kinase) assay from ChemiconInternational. Upregulation can result from over expression oractivation of growth factor receptors such as ERBB2 and EGFR, or mutantras or raf proteins.

Typical methods for screening for over expression, upregulation ormutants include, but are not limited to, standard methods such asreverse-transcriptase polymerase chain reaction (RT-PCR) or in-situhybridisation.

In screening by RT-PCR, the level of mRNA for the aforementionedproteins in the tumour is assessed by creating a cDNA copy of the mRNAfollowed by amplification of the cDNA by PCR. Methods of PCRamplification, the selection of primers, and conditions foramplification, are known to a person skilled in the art. Nucleic acidmanipulations and PCR are carried out by standard methods, as described,for example, in Ausubel, F. M. et al., eds., Current Protocols inMolecular Biology, 2004 (John Wiley & Sons Inc.); Innis, M. A. et-al.,eds., PCR Protocols: A Guide to Methods and Applications, 1990 (AcademicPress). Reactions and manipulations involving nucleic acid techniquesare also described in Sambrook et al., Molecular Cloning: A LaboratoryManual, 3rd edition, 2001 (Cold Spring Harbor Laboratory Press).Alternatively, a commercially available kit for RT-PCR (e.g., RocheMolecular Biochemicals) may be used, or methodology as set forth in U.S.Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659, 5,272,057,5,882,864, and 6,218,529.

An example of an in-situ hybridisation technique would be fluorescencein situ hybridisation (FISH) (see, e.g., Angerer, 1987, Meth. Enzymol.,Vol. 152, p. 649).

Generally, in situ hybridization comprises the following major steps:(1) fixation of tissue to be analyzed; (2) prehybridization treatment ofthe sample to increase accessibility of target nucleic acid, and toreduce nonspecific binding; (3) hybridization of the mixture of nucleicacids to the nucleic acid in the biological structure or tissue; (4)post-hybridization washes to remove nucleic acid fragments not bound inthe hybridization, and (5) detection of the hybridized nucleic acidfragments. The probes used in such applications are typically labeled,for example, with radioisotopes or fluorescent reporters. Preferredprobes are sufficiently long, for example, from about 50, 100, or 200nucleotides to about 1000 or more nucleotides, in order to enablespecific hybridization with the target nucleic acid(s) under stringentconditions. Standard methods for carrying out FISH are described, forexample, in Ausubel, F. M. et al., eds., Current Protocols in MolecularBiology, 2004 (John Wiley & Sons Inc.); Bartlett, John M. S.,“Fluorescence In Situ Hybridization Technical Overview,” in: MolecularDiagnosis of Cancer, Methods and Protocols, 2nd ed. (Series: Methods inMolecular Medicine), March 2004, pp. 77-88 (ISBN: 1-59259-760-2).

Alternatively, the protein products expressed from the mRNAs may beassayed by immunohistochemistry of tumour sections, solid phaseimmunoassay with microtiter plates, Western blotting, 2-dimensionalSDS-polyacrylamide gel electrophoresis, ELISA, and other methods knownin the art for detection of specific proteins. Detection methods wouldinclude the use of site specific antibodies, such as, phospho raf,phospho ERK, phospho MEK, or phosphotyrosine. In addition to tumourbiopsies, other samples which could be utilised include pleural fluid,peritoneal fluid, urine, stool biopsies, sputum, blood (isolation andenrichment of shed tumour cells).

In addition, mutant forms of raf, EGFR or ras can be identified bydirect sequencing of, for example, tumour biopsies using PCR and methodsto sequence PCR products directly, for example, using methods asdescribed herein. These and other well-known techniques for detection ofthe over expression, activation, or mutations may be used.

Also, abnormal levels of proteins such as raf, ras and EGFR can bemeasured using standard enzyme assays, for example for raf those assaysdescribed herein.

Alternative methods for the measurement of the over expression oractivation of FGFR, Tie, VEGFR or Eph kinases, in particular VEGFRincluding the isoforms thereof, include the measurement of microvesseldensity. This can be measured, for example, using methods described byOrre and Rogers, 1999, Int. J. Cancer, Vol. 84(2), pp. 101-108.

Assay methods also include the use of markers; for example, in the caseof VEGFR, markers include CD31, CD34 and CD105 (Mineo et al., 2004, J.Clin. Pathol., Vol. 57(6), pp. 591-597).

Treatment

The term “treatment,” as used herein in the context of treating adisease or disorder, pertains generally to treatment and therapy,whether of a human or an animal (e.g., in veterinary applications), inwhich some desired therapeutic effect is achieved, for example, theinhibition of the progress of the disease or disorder, and includes areduction in the rate of progress, a halt in the rate of progress,alleviatiation of symptoms of the disease or disorder, amelioration ofthe disease or disorder, and cure of the disease or disorder. Treatmentas a prophylactic measure (i.e., prophylaxis) is also included. Forexample, use with patients who have not yet developed the disease ordisorder, but who are at risk of developing the disease or disorder, isencompassed by the term “treatment.”

For example, treatment includes the prophylaxis of cancer, reducing theincidence of cancer, alleviating the symptoms of cancer, etc.

The term “therapeutically-effective amount,” as used herein, pertains tothat amount of a compound, or a material, composition or dosage formcomprising a compound, which is effective for producing some desiredtherapeutic effect, commensurate with a reasonable benefit/risk ratio,when administered in accordance with a desired treatment regimen.

Combination Therapies

The term “treatment” includes combination treatments and therapies, inwhich two or more treatments or therapies are combined, for example,sequentially or simultaneously. For example, the compounds describedherein may also be used in combination therapies, e.g., in conjunctionwith other agents, for example, cytotoxic agents, anticancer agents,etc. Examples of treatments and therapies include, but are not limitedto, chemotherapy (the administration of active agents, including, e.g.,drugs, antibodies (e.g., as in immunotherapy), prodrugs (e.g., as inphotodynamic therapy, GDEPT, ADEPT, etc.); surgery; radiation therapy;photodynamic therapy; gene therapy; and controlled diets.

For example, it may be beneficial to combine treatment with a compoundas described herein with one or more other (e.g., 1, 2, 3, 4) agents ortherapies that regulates cell growth or survival or differentiation viaa different mechanism, thus treating several characteristic features ofcancer development.

One aspect of the present invention pertains to a compound as describedherein, in combination with one or more additional therapeutic agents,as described below.

Examples of additional therapeutic agents that may be administeredtogether (whether concurrently or at different time intervals) with thecompounds described herein include:

-   -   (a) topoisomerase I inhibitors;    -   (b) antimetabolites;    -   (c) tubulin targeting agents;    -   (d) DNA binder and topoisomerase II inhibitors;    -   (e) alkylating agents;    -   (f) monoclonal antibodies;    -   (g) anti-hormones;    -   (h) signal transduction inhibitors;    -   (i) proteasome inhibitors;    -   (j) DNA methyl transferases;    -   (k) cytokines and retinoids.

The particular combination would be at the discretion of the physicianwho would select dosages using his common general knowledge and dosingregimens known to a skilled practitioner.

The agents (i.e., the compound described herein, plus one or more otheragents) may be administered simultaneously or sequentially, and may beadministered in individually varying dose schedules and via differentroutes. For example, when administered sequentially, the agents can beadministered at closely spaced intervals (e.g., over a period of 5-10minutes) or at longer intervals (e.g., 1, 2, 3, 4 or more hours apart,or even longer periods apart where required), the precise dosage regimenbeing commensurate with the properties of the therapeutic agent(s).

The agents (i.e., the compound described here, plus one or more otheragents) may be formulated together in a single dosage form, oralternatively, the individual agents may be formulated separately andpresented together in the form of a kit, optionally with instructionsfor their use.

Other Uses

The PDP8 compounds described herein may also be used as cell cultureadditives to inhibit RAF (e.g., B-RAF) function, e.g., to inhibit cellproliferation, etc.

The PDP8 compounds described herein may also be used as part of an invitro assay, for example, in order to determine whether a candidate hostis likely to benefit from treatment with the compound in question.

The PDP8 compounds described herein may also be used as a standard, forexample, in an assay, in order to identify other compounds, other RAF(e.g., B-RAF) function inhibitors, other anti-proliferative agents,other anti-cancer agents, etc.

Kits

One aspect of the invention pertains to a kit comprising (a) a PDP8compound as described herein, or a composition comprising a PDP8compound as described herein, e.g., preferably provided in a suitablecontainer and/or with suitable packaging; and (b) instructions for use,e.g., written instructions on how to administer the compound orcomposition.

The written instructions may also include a list of indications forwhich the active ingredient is a suitable treatment.

Routes of Administration

The PDP8 compound or pharmaceutical composition comprising the PDP8compound may be administered to a subject by any convenient route ofadministration, whether systemically/peripherally or topically (i.e., atthe site of desired action).

Routes of administration include, but are not limited to, oral (e.g., byingestion); buccal; sublingual; transdermal (including, e.g., by apatch, plaster, etc.); transmucosal (including, e.g., by a patch,plaster, etc.); intranasal (e.g., by nasal spray); ocular (e.g., byeyedrops); pulmonary (e.g., by inhalation or insufflation therapy using,e.g., via an aerosol, e.g., through the mouth or nose); rectal (e.g., bysuppository or enema); vaginal (e.g., by pessary); parenteral, forexample, by injection, including subcutaneous, intradermal,intramuscular, intravenous, intraarterial, intracardiac, intrathecal,intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal,intratracheal, subcuticular, intraarticular, subarachnoid, andintrasternal; by implant of a depot or reservoir, for example,subcutaneously or intramuscularly.

The Subject/Patient

The subject/patient may be a chordate, a vertebrate, a mammal, aplacental mammal, a marsupial (e.g., kangaroo, wombat), a rodent (e.g.,a guinea pig, a hamster, a rat, a mouse), murine (e.g., a mouse), alagomorph (e.g., a rabbit), avian (e.g., a bird), canine (e.g., a dog),feline (e.g., a cat), equine (e.g., a horse), porcine (e.g., a pig),ovine (e.g., a sheep), bovine (e.g., a cow), a primate, simian (e.g., amonkey or ape), a monkey (e.g., marmoset, baboon), an ape (e.g.,gorilla, chimpanzee, orangutang, gibbon), or a human.

Furthermore, the subject/patient may be any of its forms of development,for example, a foetus.

In one preferred embodiment, the subject/patient is a human.

Formulations

While it is possible for the PDP8 compound to be administered alone, itis preferable to present it as a pharmaceutical formulation (e.g.,composition, preparation, medicament) comprising at least one PDP8compound, as described herein, together with one or more otherpharmaceutically acceptable ingredients well known to those skilled inthe art, including, but not limited to, pharmaceutically acceptablecarriers, diluents, excipients, adjuvants, fillers, buffers,preservatives, anti-oxidants, lubricants, stabilisers, solubilisers,surfactants (e.g., wetting agents), masking agents, colouring agents,flavouring agents, and sweetening agents. The formulation may furthercomprise other active agents, for example, other therapeutic orprophylactic agents.

Thus, the present invention further provides pharmaceuticalcompositions, as defined above, and methods of making a pharmaceuticalcomposition comprising admixing at least one PDP8 compound, as describedherein, together with one or more other pharmaceutically acceptableingredients well known to those skilled in the art, e.g., carriers,diluents, excipients, etc. If formulated as discrete units (e.g.,tablets, etc.), each unit contains a predetermined amount (dosage) ofthe compound.

The term “pharmaceutically acceptable,” as used herein, pertains tocompounds, ingredients, materials, compositions, dosage forms, etc.,which are, within the scope of sound medical judgment, suitable for usein contact with the tissues of the subject in question (e.g., human)without excessive toxicity, irritation, allergic response, or otherproblem or complication, commensurate with a reasonable benefit/riskratio. Each carrier, diluent, excipient, etc. must also be “acceptable”in the sense of being compatible with the other ingredients of theformulation.

Suitable carriers, diluents, excipients, etc. can be found in standardpharmaceutical texts, for example, Remington's Pharmaceutical Sciences,18th edition, Mack Publishing Company, Easton, Pa., 1990; and Handbookof Pharmaceutical Excipients, 5th edition, 2005.

The formulations may be prepared by any methods well known in the art ofpharmacy. Such methods include the step of bringing into association thecompound with a carrier which constitutes one or more accessoryingredients. In general, the formulations are prepared by uniformly andintimately bringing into association the compound with carriers (e.g.,liquid carriers, finely divided solid carrier, etc.), and then shapingthe product, if necessary.

The formulation may be prepared to provide for rapid or slow release;immediate, delayed, timed, or sustained release; or a combinationthereof.

Formulations may suitably be in the form of liquids, solutions (e.g.,aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous),emulsions (e.g., oil-in-water, water-in-oil), elixirs, syrups,electuaries, mouthwashes, drops, tablets (including, e.g., coatedtablets), granules, powders, losenges, pastilles, capsules (including,e.g., hard and soft gelatin capsules), cachets, pills, ampoules,boluses, suppositories, pessaries, tinctures, gels, pastes, ointments,creams, lotions, oils, foams, sprays, mists, or aerosols.

Formulations may suitably be provided as a patch, adhesive plaster,bandage, dressing, or the like which is impregnated with one or morecompounds and optionally one or more other pharmaceutically acceptableingredients, including, for example, penetration, permeation, andabsorption enhancers. Formulations may also suitably be provided in theform of a depot or reservoir.

The compound may be dissolved in, suspended in, or admixed with one ormore other pharmaceutically acceptable ingredients. The compound may bepresented in a liposome or other microparticulate which is designed totarget the compound, for example, to blood components or one or moreorgans.

Formulations suitable for oral administration (e.g., by ingestion)include liquids, solutions (e.g., aqueous, non-aqueous), suspensions(e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water,water-in-oil), elixirs, syrups, electuaries, tablets, granules, powders,capsules, cachets, pills, ampoules, boluses.

Formulations suitable for buccal administration include mouthwashes,losenges, pastilles, as well as patches, adhesive plasters, depots, andreservoirs. Losenges typically comprise the compound in a flavoredbasis, usually sucrose and acacia or tragacanth. Pastilles typicallycomprise the compound in an inert matrix, such as gelatin and glycerin,or sucrose and acacia. Mouthwashes typically comprise the compound in asuitable liquid carrier.

Formulations suitable for sublingual administration include tablets,losenges, pastilles, capsules, and pills.

Formulations suitable for oral transmucosal administration includeliquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g.,aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil),mouthwashes, losenges, pastilles, as well as patches, adhesive plasters,depots, and reservoirs.

Formulations suitable for non-oral transmucosal administration includeliquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g.,aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil),suppositories, pessaries, gels, pastes, ointments, creams, lotions,oils, as well as patches, adhesive plasters, depots, and reservoirs.

Formulations suitable for transdermal administration include gels,pastes, ointments, creams, lotions, and oils, as well as patches,adhesive plasters, bandages, dressings, depots, and reservoirs.

Tablets may be made by conventional means, e.g., compression ormoulding, optionally with one or more accessory ingredients. Compressedtablets may be prepared by compressing in a suitable machine thecompound in a free-flowing form such as a powder or granules, optionallymixed with one or more binders (e.g., povidone, gelatin, acacia,sorbitol, tragacanth, hydroxypropylmethyl cellulose); fillers ordiluents (e.g., lactose, microcrystalline cellulose, calcium hydrogenphosphate); lubricants (e.g., magnesium stearate, talc, silica);disintegrants (e.g., sodium starch glycolate, cross-linked povidone,cross-linked sodium carboxymethyl cellulose); surface-active ordispersing or wetting agents (e.g., sodium lauryl sulfate);preservatives (e.g., methyl p-hydroxybenzoate, propyl p-hydroxybenzoate,sorbic acid); flavours, flavour enhancing agents, and sweeteners.Moulded tablets may be made by moulding in a suitable machine a mixtureof the powdered compound moistened with an inert liquid diluent. Thetablets may optionally be coated or scored and may be formulated so asto provide slow or controlled release of the compound therein using, forexample, hydroxypropylmethyl cellulose in varying proportions to providethe desired release profile. Tablets may optionally be provided with acoating, for example, to affect release, for example an enteric coating,to provide release in parts of the gut other than the stomach.

Ointments are typically prepared from the compound and a paraffinic or awater-miscible ointment base.

Creams are typically prepared from the compound and an oil-in-watercream base. If desired, the aqueous phase of the cream base may include,for example, at least about 30% w/w of a polyhydric alcohol, i.e., analcohol having two or more hydroxyl groups such as propylene glycol,butane-1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycoland mixtures thereof. The topical formulations may desirably include acompound which enhances absorption or penetration of the compoundthrough the skin or other affected areas. Examples of such dermalpenetration enhancers include dimethylsulfoxide and related analogues.

Emulsions are typically prepared from the compound and an oily phase,which may optionally comprise merely an emulsifier (otherwise known asan emulgent), or it may comprises a mixture of at least one emulsifierwith a fat or an oil or with both a fat and an oil. Preferably, ahydrophilic emulsifier is included together with a lipophilic emulsifierwhich acts as a stabiliser. It is also preferred to include both an oiland a fat. Together, the emulsifier(s) with or without stabiliser(s)make up the so-called emulsifying wax, and the wax together with the oiland/or fat make up the so-called emulsifying ointment base which formsthe oily dispersed phase of the cream formulations.

Suitable emulgents and emulsion stabilisers include Tween 60, Span 80,cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodiumlauryl sulfate. The choice of suitable oils or fats for the formulationis based on achieving the desired cosmetic properties, since thesolubility of the compound in most oils likely to be used inpharmaceutical emulsion formulations may be very low. Thus the creamshould preferably be a non-greasy, non-staining and washable productwith suitable consistency to avoid leakage from tubes or othercontainers. Straight or branched chain, mono- or dibasic alkyl esterssuch as di-isoadipate, isocetyl stearate, propylene glycol diester ofcoconut fatty acids, isopropyl myristate, decyl oleate, isopropylpalmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branchedchain esters known as Crodamol CAP may be used, the last three beingpreferred esters. These may be used alone or in combination depending onthe properties required. Alternatively, high melting point lipids suchas white soft paraffin and/or liquid paraffin or other mineral oils canbe used.

Formulations suitable for intranasal administration, where the carrieris a liquid, include, for example, nasal spray, nasal drops, or byaerosol administration by nebuliser, include aqueous or oily solutionsof the compound.

Formulations suitable for intranasal administration, where the carrieris a solid, include, for example, those presented as a coarse powderhaving a particle size, for example, in the range of about 20 to about500 microns which is administered in the manner in which snuff is taken,i.e., by rapid inhalation through the nasal passage from a container ofthe powder held close up to the nose.

Formulations suitable for pulmonary administration (e.g., by inhalationor insufflation therapy) include those presented as an aerosol sprayfrom a pressurised pack, with the use of a suitable propellant, such asdichlorodifluoromethane, trichlorofluoromethane,dichoro-tetrafluoroethane, carbon dioxide, or other suitable gases.

Formulations suitable for ocular administration include eye dropswherein the compound is dissolved or suspended in a suitable carrier,especially an aqueous solvent for the compound.

Formulations suitable for rectal administration may be presented as asuppository with a suitable base comprising, for example, natural orhardened oils, waxes, fats, semi-liquid or liquid polyols, for example,cocoa butter or a salicylate; or as a solution or suspension fortreatment by enema.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the compound, such carriers as are known inthe art to be appropriate.

Formulations suitable for parenteral administration (e.g., byinjection), include aqueous or non-aqueous, isotonic, pyrogen-free,sterile liquids (e.g., solutions, suspensions), in which the compound isdissolved, suspended, or otherwise provided (e.g., in a liposome orother microparticulate). Such liquids may additional contain otherpharmaceutically acceptable ingredients, such as anti-oxidants, buffers,preservatives, stabilisers, bacteriostats, suspending agents, thickeningagents, and solutes which render the formulation isotonic with the blood(or other relevant bodily fluid) of the intended recipient. Examples ofexcipients include, for example, water, alcohols, polyols, glycerol,vegetable oils, and the like. Examples of suitable isotonic carriers foruse in such formulations include Sodium Chloride Injection, RingersSolution, or Lactated Ringer's Injection. Typically, the concentrationof the compound in the liquid is from about 1 ng/ml to about 10 μg/ml,for example from about 10 ng/ml to about 1 μg/ml. The formulations maybe presented in unit-dose or multi-dose sealed containers, for example,ampoules and vials, and may be stored in a freeze-dried (lyophilised)condition requiring only the addition of the sterile liquid carrier, forexample water for injections, immediately prior to use. Extemporaneousinjection solutions and suspensions may be prepared from sterilepowders, granules, and tablets.

Dosage

It will be appreciated by one of skill in the art that appropriatedosages of the PDP8 compounds, and compositions comprising the PDP8compounds, can vary from patient to patient. Determining the optimaldosage will generally involve the balancing of the level of therapeuticbenefit against any risk or deleterious side effects. The selecteddosage level will depend on a variety of factors including, but notlimited to, the activity of the particular PDP8 compound, the route ofadministration, the time of administration, the rate of excretion of thePDP8 compound, the duration of the treatment, other drugs, compounds,and/or materials used in combination, the severity of the disease ordisorder, and the species, sex, age, weight, condition, general health,and prior medical history of the patient. The amount of PDP8 compoundand route of administration will ultimately be at the discretion of thephysician, veterinarian, or clinician, although generally the dosagewill be selected to achieve local concentrations at the site of actionwhich achieve the desired effect without causing substantial harmful ordeleterious side-effects.

Administration can be effected in one dose, continuously orintermittently (e.g., in divided doses at appropriate intervals)throughout the course of treatment. Methods of determining the mosteffective means and dosage of administration are well known to those ofskill in the art and will vary with the formulation used for therapy,the purpose of the therapy, the target cell(s) being treated, and thesubject being treated. Single or multiple administrations can be carriedout with the dose level and pattern being selected by the treatingphysician, veterinarian, or clinician.

In general, a suitable dose of the PDP8 compound is in the range ofabout 10 μg to about 250 mg (more typically about 100 μg to about 25 mg)per kilogram body weight of the subject per day. Where the compound is asalt, an ester, an amide, a prodrug, or the like, the amountadministered is calculated on the basis of the parent compound and sothe actual weight to be used is increased proportionately.

Examples

The following examples are provided solely to illustrate the presentinvention and are not intended to limit the scope of the invention, asdescribed herein.

Chemical Synthesis

Several methods for the chemical synthesis of compounds of the presentinvention are described herein. These and/or other well known methodsmay be modified and/or adapted in known ways in order to facilitate thesynthesis of additional compounds within the scope of the presentinvention.

Descriptions of general laboratory methods and procedures, useful forthe preparation of the compounds described herein, are provided inVogel's Textbook of Practical Organic Chemistry, 5th Edition, 1989,(Editors: Furniss, B. S., Hannaford, A. J., Smith, P. W. G., Tatchell,A. R.) (published by Longmann, UK).

Methods for the synthesis of pyridine compounds in particular aredescribed in Heterocyclic Chemistry, 3rd Edition, 1998, Joule, J. A,Mills, R. and Smith, G. F. (published by Chapman & Hall, UK).

Many of the compounds described herein can be prepared via a keyintermediate (2), conveniently substituted on the aromatic ring. Thisintermediate can be prepared from commercially available startingmaterial, 4-chloro-3-nitro-pyridin-2-amine, (1), and substitutedamino-phenols. Compounds 2 are then protected selectively at the aminogroup, for example as a Boc carbamate or trifluoroacetamide, to affordintermediates, (3). The intermediates, (3), can also be obtaineddirectly from 4-chloro-3-nitro-pyridin-2-amine, (1), and N-Boc-protectedamino-phenols. The nitro group of the protected intermediate, (3), maybe reduced to an amino group with Pd/C and ammonium formate or hydrogen,to another key diamino intermediate (4). An example of such a method isillustrated in the following Scheme 1.

Note that compounds with substituted or unsubstituted phenyl groups havebeen synthesised and are described herein. The following Schemes areillustrated using unsubstituted phenyl or specifically substitutedphenyl, but it should be understood that these methods are also suitablefor the preparation of compounds with substituted (or differentlysubstituted) phenyl rings.

Pyridopyrazinones can be obtained from intermediate 4 by reaction withethyl glyoxylate, ethyl pyruvate or similar α-ketoesters. Both isomers 5and 6 can be obtained from the reaction of 4 with ethyl glyoxalate.Similarly, two isomers (7 and 8) can be obtained from the reaction of 4with ethyl pyruvate (R═-Me), ethyltrifluoropyruvate (R═—CF₃), ethyl3-bromo-2-oxopropanoate (R═—CH₂Br), or other optionally substitutedalkyl 2-oxo esters. Amino-pyridopyrazinones 9 and 10 can be obtainedfrom intermediate 4 by reaction with ethyl 2-ethoxy-2-iminoacetate. Theratio of the two isomers can be influenced by the choice of solvents, sothat one is obtained preferentially (Scheme 2).

Deprotection of the protecting group (PG) with TFA or tetrabutylammonium fluoride (for Boc protecting group) or ammonia (fortrifluoroacetamide) produces the common intermediates 11-16 (Scheme 3).

Pyridopyrazines 18 can be obtained from intermediate 4 by reaction withglyoxal or 1,4-dioxane-2,3-diol followed by deprotection (Scheme 4).

Pyridopyrazin-diones 20 can be obtained from intermediate 4 by reactionwith diethyloxalate or oxalyl chloride followed by deprotection (Scheme5).

Amino-pyridopyrazines 25 and 26 can be obtained from intermediates 5 and6. The carbonyl group of the pyrazinone can be converted to thechloropyrazine intermediates 21 or 22 with POCl₃ or NCS/PPh₃, then toaminopyrazines 23 or 24 using ammonia or primary or secondary amine.Deprotection affords the common intermediates 25 or 26 (Scheme 6).

These common intermediates (11-16, 18, 20, 25, 26) may then be used toprepare a range of compounds with different linker groups, L, anddifferent terminal groups, A. For example, the key intermediate 11 canbe reacted with activated carboxylic acids or acid chlorides to affordamides (NHCO) or with isocyanates or with activated carbamates to affordureas (NHCONH). Isocyanates can also be formed in situ by reaction ofcarboxylic acids with, for example, DPPA (diphenylphosphoryl azide) andCurtius rearrangement of the corresponding azide upon heating. The keyintermediate 11 can also react with isothiocyanates to afford thioureas(NHCSNH) and with sulfonyl chlorides to afford sulfonamides (SO₂NH).Examples of such methods are illustrated in the following scheme (Scheme7).

Alternatively, the amino position of the common intermediate 11 can beactivated by reaction, for example, with phenyl chloroformate. Theactivated carbamate so formed can then be reacted with aromatic aminesto afford the corresponding ureas, as illustrated in Scheme 8.

An alternative strategy is to perform the reactions described in Schemes7 and 8 (formation of urea or amide) on the nitro-amino intermediate 2prior to cyclisation. Similarly, amino phenols can react withisocyanates to form the intermediate 30, which is then coupled with 1 toafford 28. Such an approach is exemplified for the urea linker in Scheme9. Similar methods can be used for compounds with other linkers.

Compounds with reverse amide linker can be obtained from reaction ofstarting material 1 with hydroxyl-benzoic acids. For example, 1 canreact with methyl 3-hydroxybenzoate to form intermediate 31. Thisintermediate can be reduced to diamino intermediate 32 and cyclised toone of the scaffolds described in Schemes 2, 4-6 then reacted with arylor heteroaryl amine to afford the final product. For exemplification isformation of pyridopyrazinone 34 via the intermediate ester 33.Alternatively, intermediate 31 can be reacted with aryl amine to afford35, reduced to diamine 36 and cyclised to the same product 34 (Scheme10).

Compounds with other linkers between the hinge-binding bicyclic systemand middle ring can be obtained by reacting the starting material 1,with for example mercaptoanilines, aminoanilines or mercaptobenzoicesters, as exemplified in Scheme 11. The intermediates thus obtained canbe converted further to inhibitors containing the same scaffoldsdescribed for the O-linker compounds using methods similar to those inSchemes 1-10.

Chemical Synthesis

All starting materials, reagents and solvents for reactions were reagentgrade and used as purchased. Chromatography solvents were HPLC grade andwere used without further purification. Reactions were monitored by thinlayer chromatography (TLC) analysis using Merck silica gel 60 F-254 thinlayer plates. Flash column chromatography was carried out on Mercksilica gel 60 (0.015-0.040 mm) or in disposable Isolute Flash Si and SiII silica gel columns. Preparative TLC was performed on eitherMacherey-Nagel [809 023] pre-coated TLC plates SIL G-25 UV₂₅₄ orAnaltech [2015] pre-coated preparative TLC plates, 2000 μm with UV₂₅₄.LCMS analyses were performed on a Micromass LCT/Water's Alliance 2795HPLC system with a Discovery 5 μm, C18, 50 mm×4.6 mm i.d. column fromSupelco at a temperature of 22° C. using the following solvent systems:Solvent A: Methanol; Solvent B: 0.1% formic acid in water at a flow rateof 1 mL/min. Gradient starting with 10% A/90% B from 0-0.5 minutes then10% A/90% B to 90% A/10% B from 0.5 minutes to 6.5 minutes andcontinuing at 90% A/10% B up to 10 minutes. From 10-10.5 minutes thegradient reverted back to 10% A/90% where the concentrations remaineduntil 12 minutes. UV detection was at 254 nm and ionisation was positiveor negative ion electrospray. Molecular weight scan range is 50-1000.Samples were supplied as 1 mg/mL in DMSO or methanol with 3 μL injectedon a partial loop fill. NMR spectra were recorded in DMSO-d₆ on a BrukerAdvance 500 MHz spectrometer.

(I) Coupling of 2-Amino-3-Nitro-4-Chloropyridine with PhenolatesSynthesis 1 Tert-butyl 4-(2-amino-3-nitropyridin-4-yloxy)phenylcarbamate

Method A1: Tert-butyl 4-hydroxyphenylcarbamate (3.63 g, 17.4 mmol) wasdissolved in dry DMF (150 mL). Potassium tert-butoxide (2.62 g, 23.4mmol) was added and the stirring was continued for 30 minutes at roomtemperature. 4-Chloro-3-nitropyridin-2-amine (3.0 g, 17.3 mmol) wasadded as a solid in one portion and the reaction mixture wassubsequently heated at 85° C. for 4 hours. The reaction mixture wascooled, diluted with ethyl acetate (800 ml) and washed with water (1×800ml) and brine (2×800 ml). The organic layer was dried with magnesiumsulphate and evaporated. The crude was chromatographed over silica(eluant ethyl acetate:cyclohexane 1:2) to yield 4.0 g (63 yield) oftert-butyl 4-(2-amino-3-nitropyridin-4-yloxy)phenylcarbamate.

¹H-NMR (CDCl₃), δ (ppm), J (Hz): ¹H-NMR, δ (ppm), J (Hz): 1.54 (9H),6.04 (d, 1H, J=7.4 Hz), 6.15 (bs, 2H), 7.06 (d, 2H, J=8.3 Hz), 7.44 (d,2H, J=8.3 Hz), 7.96 (d, 1H, J=7.4 Hz). LC-MS (m/z): 347 (M+H, 100).

Synthesis 2 Tert-butyl4-(2-amino-3-nitropyridin-4-yloxy)naphthalen-1-ylcarbamate

Method A1 was used with tert-butyl 4-hydroxynaphthalen-1-ylcarbamate(3.9 g, 15 mmol) to yield tert-butyl4-(2-amino-3-nitropyridin-4-yloxy)naphthalen-1-ylcarbamate (5.4 g, 90%yield) upon recrystallization from dichloromethane.

¹H-NMR (CDCl₃), δ (ppm), J (Hz): 1.58 (s, 9H), 5.92 (d, 1H, J=5.8 Hz),6.21 (s, 1H), 7.25 (d, 1H, J=8.3 Hz), 7.56 (t, 1H, J=8.1 Hz), 7.62 (t,1H, J=8.3 Hz), 7.88 (d, 1H, J=5.8 Hz), 7.93 (s, 1H), 7.95 (d, 1H, J=8.5Hz), 8.00 (d, 1H, J=8.3 Hz), LC-MS: m/z 397 (M+H, 100).

Synthesis 3 4-(4-amino-3-(methylthio)phenoxy)-3-nitropyridin-2-amine

Method A2 Sodium hydride (148 mg) was added to dry DMSO (5.5 mL) and themixture was stirred at RT for 20 minutes under Ar atmosphere.4-amino-3-(methylthio)phenol (573 mg, 3.7 mmol) was added thereto, andthe mixture stirred for 10 more minutes. Next,4-Chloro-3-nitropyridin-2-amine (3.7 mmol) was added, and the mixturewas heated to 100° C. and stirred for 3 hours. After cooling down, waterwas added, and the mixture extracted three times with EtOAc. Thecombined organic layers were washed first with a saturated aqueoussodium hydrogen carbonate solution then water, dried over MgSO₄ andevaporated to afford the title compound (657 mg, 61%) was obtained afterpurification by chromatography on silica gel (EtOAc-DCM, 1:1) as a redbrown solid (R_(f) 0.56, EtOAc-DCM, 1:1).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.36 (s, 3H, CH₃); 5.18 (s, 2H,NH_(2, Ph)), 5.92 (d, 1H, H_(Py) J=5.8 Hz), 6.75 (dd, 1H, H_(Ph) J=8.6Hz and J=2.1 Hz), 6.81 (dd, 1H, H_(Ph) J=8.7 and J=2.6 Hz), 6.98 (d, 1H,H_(Ph) J=2.6 Hz), 7.07 (bs, 2H, NH_(2, Py)), 7.95 (d, 1H, H_(Py) J=5.7Hz). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 15.6, 99.8, 114.7, 119.7,120.7, 121.4, 121.5, 143.2, 145.1, 152.8, 153.6, 159.9. LC-MS (m/z): 293(M+H, 100), rt=5.87 min.

Synthesis 4 4-amino-3-(methylthio)phenol

Method C4: A suspension of iron powder (220 mg, 4 mmol), NH₄Cl (310 mg,5.8 mmol) in a mixture EtOH/H₂O (4 mL/1.2 mL) was heated to reflux for10 minutes. 3-(methylthio)-4-nitrophenol (185 mg, 1 mmol) was added andthe mixture stirred for 5 hours. After cooling to RT, the dark slurrywas filtered over celite and washed with MeOH. After removing thesolvent, EtOAc was added and the mixture filtered once again. Thefiltrate was washed successively with water and brine, and then driedover MgSO₄. Removal of the solvent under vacuum provided the titlecompound as a green-grey powder (80 mg, 53% yield).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.29 (s, 3H, H_(Me)), 4.48 (bs, 2H,NH₂), 6.44 (d, 1H, H_(arom), J=8.5 Hz), 6.54 (d, 1H, H_(arom) J=8.5 Hz),6.61 (s, 1H, H_(arom)), 8.58 (bs, 1H, OH). ¹³C-NMR (DMSO-d₆), δ (ppm), J(Hz): 15.9, 114.7, 115.4, 116.5, 120.1, 139.5, 148.7. GC-MS (m/z):155.09.

Synthesis 5 3-(methylthio)-4-nitrophenol

To a solution of 3-fluoro-4-nitrophenol (2 g, 12.7 mmol) in dry DMF (67mL) were added by aliquots 2 equivalents of sodium thiomethoxide (1.78g, 25.5 mmol) followed by 3 equivalents of potassium carbonate (5.27 g,38.2 mmol). The mixture was stirred at RT for 23 hours and then water(100 mL) was added. The mixture was extracted with EtOAc, and thecombined organic layers washed successively with water (60 mL) and brine(60 mL) and then dried over MgSO₄. The solvent was evaporated undervacuum to provide the title compound (2.12 g, 90%) as a yellow powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.44 (s, 3H, H_(Me)), 6.72 (d, 1H,H_(arom), J=9.0 Hz), 6.79 (s, 1H, H_(arom)), 8.19 (d, 1H, H_(arom),J=9.1 Hz), 11.20 (bs, 1H, OH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 15.2,111.3, 112.0, 128.7, 136.7, 142.0, 162.9.

Synthesis 6 4-(4-Amino-3-fluorophenoxy)-3-nitropyridin-2-amine

Method A2 was used with 4-amino-3-fluorophenol (1.00 g, 7.9 mmol) togive 1.8 g (86% yield) of4-(4-amino-3-fluorophenoxy)-3-nitropyridin-2-amine as a dark solid.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.17 (bs, 2H), 5.94 (d, 1H, J=5.7Hz), 6.75-6.84 (m, 2H), 6.97 (d, 1H, J=11.7 Hz) 7.09 (bs, 1H), 7.96 (d,1H, J=5.7 Hz). LC-MS (m/z): 235 (M+H, 100).

Synthesis 74-(4-N-(tert-Butoxycarbonyl)amino-3-fluorophenoxy)-3-nitro-2-amino-pyridine

Method A1 was used with 4-N-Boc-amino-3-fluorophenol (1.2 g, 5.4 mmol)to afford the title compound as a glassy yellow solid (1.9 g, 96%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu), 6.08 (d, 1H,J=5.5, H_(Py)), 7.01 (m, 1H, H_(arom)), 7.18 (br s, 2H, NH₂), 7.22 (m,1H, H_(arom)), 7.67 (m, 1H, H_(arom)), 8.04 (d, 1H, J=5.5, H_(Py)), 9.03(s, 1H, NH_(Boc)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.0, 79.5,100.7, 108.8 (d, J_(FC)=23.1), 116.2 (d, J_(FC)=3.1), 121.7, 124.3 (d,J_(FC)=12.2), 125.4, 149.4 (d, J_(FC)=10.1), 153.0, 153.3, 153.9, 154.1(d, J_(FC)=249), 158.6; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −120.7; LC-MS (m/z):365.0 (M+H, 100).

Synthesis 8 Tert-butyl 2-fluoro-4-hydroxyphenylcarbamate

Method B: 4-amino-3-fluorophenol (10.61 g, 83.5 mmol) was added to amolten mixture of Boc₂O (18.29 g, 83.8 mmol) and InCl₃ (188 mg, 0.85mmol) at 35° C. The black mixture was stirred at 35° C. for 2 h, duringwhich time it turned into a thick black oil. The mixture was thendiluted with EtOAc (200 mL) and H₂O (200 mL) and stirring was continuedfor 10 min. The layers were separated and the organic layer was washedwith H₂O (3×200 mL), dried (MgSO₄), filtered and concentrated todryness. The resulting black oil was redissolved in CH₂Cl₂ (50 mL) andloaded onto a silica gel column. Elution with 57% EtOAc in CH₂Cl₂furnished the title compound as a light yellow, crystalline solid.Yield: 16.7 g (90%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu), 6.08 (d, 1H,J=5.5, H_(Py)), 7.01 (m, 1H, H_(arom)), 7.18 (br s, 2H, NH₂), 7.22 (m,1H, H_(arom)), 7.67 (m, 1H, H_(arom)), 8.04 (d, 1H, J=5.5, _(Py)rH),9.03 (s, 1H, NH_(Boc)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.0, 78.6,102.7 d, (J_(FC)=22.2), 110.8 (d, J_(FC)=2.7), 117.1 (d, J_(FC)=12.6),127.2, 153.7, 155.5 (d, J_(FC)=11.3), 156.1 (d, J_(FC)=246); ¹⁹F-NMR(DMSO-d₆), δ (ppm): −121.6; LC-MS (m/z): 172.0 (M+H, 100).

Synthesis 9 tert-butyl-4-hydroxy-3-fluorophenylcarbamate

Using Method B with 4-amino-2-fluorophenol (1.6 g, 12.7 mmol), the titlecompound (1.26 g, 44%) was obtained after 1 hour, and purified usingBiotage (EtOAc-DCM: 1-1) to give a pale pink powder. (Rf 0.86,EtOAc-DCM, 1-1).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu); 6.82 (t, 1H,H_(arom), J=9.2 Hz), 6.99 (d, 1H, H_(arom), J=8.1 Hz), 7.29 (d, 1H,H_(arom), J=13.5 Hz), 9.18 (s, 1H, OH), 9.36 (s, 1H, NH_(carbamate)).¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.9, 79.9, 107.9, 115.4, 118.5,132.6, 140.3, 150.4, 152.3. ¹⁹F-NMR (δ, ppm, DMSO-d6): −134.62.

Synthesis 10 tert-butyl4-(2-amino-3-nitropyridin-4-yloxy)-3-fluorophenylcarbamate

Using Method A1 with tert-butyl-4-hydroxy-3-fluorophenylcarbamate (1.26g, 5.5 mmol), the title compound (1.99 g, 99%) was obtained after 1 hourstirring as a yellow powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.52 (s, 9H, tert-Bu); 5.98 (d, 1H,H_(Py) J=5.7 Hz), 7.21 (s, 2H, NH₂), 7.32 (m, 2H, H_(arom)), 7.63 (m,1H, H_(arom)), 8.02 (d, 1H, H_(Py) J=5.4 Hz), 9.74 (s, 1H,NH_(carbamate)). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.0, 79.7, 99.0,106.3, 114.6, 121.0, 123.5, 133.7, 139.0, 152.6, 153.2, 153.7, 154.0,158.7. ¹⁹F-NMR (δ, ppm, DMSO-d₆): −128.76. LC-MS (m/z): 365 (M+H, 100),rt=2.58 min.

Synthesis 114-(3-N-(tert-Butoxycarbonyl)aminophenoxy)-3-nitro-2-amino-pyridine

Method A1 was used with 3-N-Boc-amino-phenol (1.2 g, 5.4 mmol) to affordthe title compound as a glassy yellow solid (1.7 g, 90%).

¹H-NMR (DMSO), δ (ppm), J (Hz): 1.46 (s, 9H, (CH₃)₃C), 5.36 (s, 2H,NH₂), 6.00 (d, 1H, H_(Pyr), J=5.7), 6.77 (d, 1H, H_(arom), J=6.9),7.32-7.36 (m, 2H, H_(arom)), 8.01 (d, 1H, H_(Pyr)), 9.56 (s, 1H, NH);LC-MS (m/z): 346.1 (M+H, 100), rt=7.10 min.

Synthesis 12 Methyl 3-(2-amino-3-nitropyridin-4-yloxy)benzoate

Method A1 was used with methyl 3-hydroxybenzoate (800 mg, 4.7 mmol) toafford the title compound (760 mg, 53% yield).

¹H-NMR (DMSO), δ (ppm), J (Hz): 3.86 (s, 3H, Me), 6.04 (d, 1H, H_(Pyr),J=6.0 Hz), 7.23 (s, 2H, NH₂), 7.52 (d, 1H, H_(arom), J=8.0 Hz),7.63-7.66 (m, 1H, H_(arom)), 7.88 (d, 1H, H_(arom), J=8.0 Hz), 8.04 (d,1H, H_(Pyr)); LC-MS (m/z): 290 (M+H, 100).

Synthesis 13 Tert-butyl4-(2-amino-3-nitropyridin-4-ylthio)phenylcarbamate

Method A3: Dry DMSO (15 mL) was added to NaH (1.24 g of a 60% dispersionin mineral oil, 25.7 mmol) in a round bottom flask under Ar. After 5min, solid tert-butyl 4-mercaptophenylcarbamate (6.98 g, 31.0 mmol) wasadded in three portions, which led to the formation of a yellow solutionwhile effervescence occurred. After 15 min of stirring at RT,4-chloro-3-nitropyridin-2-amine (5.38 g, 31.0 mmol) at once. Theyellow/brown solution was stirred for 30 min and EtOAc (150 mL) and H₂O(400 mL) were subsequently added. The aqueous layer was extracted withEtOAc (3×100 mL) and the combined organic layers were washed once withsaturated NaHCO₃ (150 mL), dried (MgSO₄), filtered, and concentrated todryness to give the title compound as a bright yellow solid. Yield: 11.2g (quantitative).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.49 (s, 9H, tert-Bu), 5.83 (d,J=5.4, 1H, H_(Py)), 7.47 (d, J=8.7, 2H, H_(arom)), 7.64 (d, J=8.7, 2H,H_(arom)), 7.87-7.89 (m, 3H), 9.69 (s, 1H, NHBoc); ¹³C-NMR (DMSO-d₆), δ(ppm), J (Hz): 28.0, 79.6, 110.3, 119.4, 121.5, 124.8, 136.4, 141.4,152.3, 152.5, 153.6, 156.2; LC-MS: 364.0 (M+H, 100); HRMS: m/z calcd.for C₁₆H₁₃N₄O₄S [M+H⁺]: 363.11215. Found: 363.11261.

Synthesis 14 Tert-butyl 4-mercaptophenylcarbamate

Method B was used with 4-aminobenzenethiol (8.08 g, 64.5 mmol) to affordthe title compound Yield: 14.5 g (100%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu), 5.08 (s, 1H,SH), 7.17 (d, J=8.7, 2H, H_(arom)), 7.34 (d, J=8.7, 2H, H_(arom)), 9.27(s, 1H, NH_(Boc)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.1, 79.0,118.9, 123.4, 129.6, 130.7, 137.2, 140.0, 152.7.

(II) Boc Protection of Amine Synthesis 154-(4-N-(tert-Butoxycarbonyl)amino-3-thiomethyl-phenoxy)-3-nitro-2-amino-pyridine

Method B, was used with4-(4-amino-3-(methylthio)phenoxy)-3-nitropyridin-2-amine (2 g, 6.8mmol). The title compound (2.42 g, 90%) was obtained after purificationby chromatography on silica gel (EtOAc-DCM: 1-1, then EtOAc-MeOH: 95-5)as a powder (R_(f) 0.33, EtOAc-MeOH, 95:5).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu); 2.81 (s, 3H,CH₃); 6.07 (d, 1H, H_(Py) J=5.6 Hz), 7.19 (m, 1H, H_(arom)), 7.35 (m,1H, H_(arom)), 7.53 (d, 1H, H_(arom), J=2.8 Hz), 7.55 (d, 1H, H_(arom),J=8.7 Hz), 8.01 (m, 1H, H_(arom)), 8.05 (d, 1H, H_(Py) J=5.6 Hz), 9.32(s, 1H, NH_(carbamate)). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 14.8, 27.9,78.8, 103.6, 115.9, 117.3, 121.7, 124.6, 127.5, 132.1, 137.1, 146.0,148.5, 151.7, 153.4. LC-MS (m/z): 393 (M+H, 100), rt=7.64 min.

(III) Reduction of Nitro Group En-Route to Common IntermediatesAccording to Scheme 1 Synthesis 16 Tert-butyl4-(2,3-diaminopyridin-4-yloxy)phenylcarbamate

Method C1: 1.56 g (4.5 mmol) of tert-butyl4-(2-amino-3-nitropyridin-4-yloxy)phenylcarbamate are dissolved in 300ml of a 1:1 ethanol:ethyl acetate mixture. The solution was mixed withH₂ was passed through a cartridge containing Pd/C in a H-cube apparatus,then was evaporated to provide 1.26 g (88% yield) of tert-butyl4-(2,3-diaminopyridin-4-yloxy)phenylcarbamate as a white foamy solid.

¹H-NMR (CDCl₃), δ (ppm), J (Hz): 1.54 (9H, s), 2.90 (4H, bs), 6.60 (1H,bs), 6.17 (d, 1H, J=5.7 Hz), 7.01 (2H, d, J=8.9 Hz), 7.38 (d, 2H, J=8.9Hz), 7.52 (d, 1H, J=5.8 Hz). LC-MS (m/z): 317 (M+H, 100).

Synthesis 17 tert-butyl4-(2,3-diaminopyridin-4-yloxy)naphthalen-1-ylcarbamate

Method C1 was used with tert-butyl4-(2-amino-3-nitropyridin-4-yloxy)naphthalen-1-ylcarbamate (3.0 g, 7.6mmol) with solvent mixture MeOH:THF 1:1 to afford the title compound inquantitative yield (2.4 g).

¹H-NMR (CDCl₃), δ (ppm), J (Hz): 1.56 (s, 9H), 6.03 (d, 1H, J=6.0 Hz),7.04 (s, 1H), 7.07 (d, 1H, J=8.2 Hz), 7.33 (d, 1H, J=6.0 Hz), 7.50 (t,1H, J=7.4 Hz), 7.57 (t, 1H, J=8.2 Hz), 7.77 (bs, 1H), 7.95 (d, 1H, J=8.2Hz), 7.98 (d, 1H, J=8.2 Hz). LC-MS (m/z): 367 (M+H, 100).

Synthesis 184-(4-N-(tert-Butoxycarbonyl)amino-3-fluorophenoxy)-2,3-diamino-pyridine

Method C2: Pd/C (1.09 g) was added to a yellow solution of4-(4-N-(tert-Butoxycarbonyl)amino-3-fluorophenoxy)-3-nitro-2-amino-pyridine(6.20 g, 17.0 mmol) in EtOAc/EtOH (90/150 mL) and the black mixture wasstirred under a hydrogen atmosphere for 5 h and filtered over Celite.The dark brown filtrate was concentrated to dryness, redissolved inCH₂Cl₂ (20 mL) and loaded onto a silicagel column. The products wereeluted with EtOAc and the fractions containing the title compound werecompound and evaporated to dryness. The orange oil was dissolved inCH₂Cl₂ and an equal amount of hexane was added. The solution wasconcentrated to dryness to give an orange foam. Yield: 4.30 g (76%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): δ=8.82 (br s, 1H, NH_(Boc)), 7.47 (t,J=8.5 Hz, 1H, H_(arom)), 7.28 (d, 1H, J=5.5 Hz, H_(Py)), 6.87 (m, 1H,H_(arom)), 6.76 (m, 1H, H_(arom)), 6.09 (d, 1H, J=5.5 Hz, H_(Py)), 5.61(s, 2H, NH₂), 4.47 (s, 2H, NH₂), 1.45 ppm (s, 9H, tert-Bu); ¹⁹F NMR (470MHz, DMSO-d₆): δ=−120.7 ppm; LC-MS (m/z): 335.3 (M+H, 100), rt=2.69 min.

Synthesis 19 Tert-butyl4-(2,3-diaminopyridin-4-yloxy)-2-(methylthio)phenylcarbamate

Using Method C4 with4-(4-N-(tert-Butoxycarbonyl)amino-3-thiomethyl-phenoxy)-3-nitro-2-amino-pyridine(12.5 g, 31.8 mmol), the title compound (2.07 g, 18%) was obtained afterpurification by chromatography on silica gel (EtOAc, then EtOAc-MeOH:95-5) as a powder (R_(f) 0.33, EtOAc-MeOH, 95:5).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.44 (s, 9H, tert-Bu); 2.39 (s, 3H,CH₃); 5.56 (bs, 2H, NH₂); 6.29 (d, 1H, H_(Py) J=6.9 Hz), 6.87 (dd, 1H,H_(arom) J=8.6 Hz, J=2.7 Hz), 7.06 (d, 1H, H_(arom) J=2.7 Hz), 7.31 (m,2H, H_(Py) J=6.8 Hz+H_(arom)), 7.56 (bs, 2H, NH_(2, Py)), 8.44 (s, 1H,NH_(carbamate)). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 14.8, 27.9, 78.8,103.6, 115.9, 117.3, 121.7, 124.6, 127.5, 132.1, 137.1, 146.0, 148.5,151.7, 153.4. LC-MS (m/z): 362 (M+H, 100), rt=3.04 min.

Synthesis 204-(3-N-(tert-Butoxycarbonyl)aminophenoxy)-2,3-diamino-pyridine

Method C2 was used with4-(3-N-(tert-butoxycarbonyl)aminophenyloxy)-2-amino-3-nitro-pyridine(2.5 g, 7.2 mmol) to afford the title compound as a brown glassy solid(2.17 g, 95%).

¹H-NMR (DMSO), δ (ppm), J (Hz): 1.45 (s, 9H, (CH₃)₃C), 4.39 (s, 2H,5-NH₂), 5.36 (s, 2H, 6-NH₂), 6.02 (d, 1H, H_(Py) J=5.6), 6.58 (d, 1H,H_(arom)J=7.9), 7.19-7.21 (m, 2H, H_(arom)), 7.25 (d, 1H, H_(Py)), 9.41(s, 1H, NH); LC-MS (m/z): 316.1 (M+H, 100), rt=4.03 min.

Synthesis 21 Methyl 3-(2,3-diaminopyridin-4-yloxy)benzoate

Method C2 was used with methyl 3-(2-amino-3-nitropyridin-4-yloxy) (760mg, 2.6 mmol), enzoate to afford the title compound (680 mg, 100%).

¹H-NMR (DMSO), δ (ppm), J (Hz): 3.83 (s, 3H, Me), 4.54 (s, 2H, NH₂),5.68 (s, 2H, NH₂), 6.12 (d, 1H, H_(Pyr), J=6.0 Hz), 7.27-7.32 (m, 1H,H_(arom)), 7.43 (d, 1H, H_(arom), J=1.5 Hz), 7.52 (t, 1H, H_(arom),J=8.0 Hz), 7.69 (d, 1H, H_(Pyr)); LC-MS (m/z): 260 (M+H, 100).

Synthesis 22 tert-butyl4-(2,3-diaminopyridin-4-yloxy)-3-fluorophenylcarbamate

Using Method C2 with tert-butyl4-(2-amino-3-nitropyridin-4-yloxy)-3-fluorophenylcarbamate (2.15 g, 5.9mmol), the title compound (1.75 g, 89%) was obtained as a brown solid.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.52 (s, 9H, tert-Bu); 4.51 (bs, 2H,NH₂), 5.59 (s, 2H, NH₂), 5.88 (d, 1H, H_(Py),5, J=4.8 Hz), 7.11 (t, 1H,J=8.8 Hz), 7.22-7.27 (m, 2H, H_(arom)), 7.56 (dd, 1H, H_(arom), J=12.2Hz J=1.6 Hz), 9.61 (s, 1H, NH_(carbamate)). ¹³C-NMR (DMSO-Cl₆), δ (ppm),J (Hz): 29.1, 80.6, 102.2, 107.6, 115.4, 119.2, 123.6, 136.6, 137.3,138.2, 149.2, 150.9, 153.7, 155.1. ¹⁹F-NMR (δ, ppm, DMSO-d6): −129.68.LC-MS (m/z): 335 (M+H, 100), rt=2.00 min.

Synthesis 23 Tert-butyl 4-(2,3-diaminopyridin-4-ylthio)phenylcarbamate

Method C3: Tert-butyl 4-(2-amino-3-nitropyridin-4-ylthio)phenylcarbamate(470 mg, 1.30 mmol) was dissolved in a mixture of EtOAc and EtOH (80mL/40 mL) and Raney nickel (a spoonful) was added. The suspension wasstirred under an H₂ atmosphere for 90 min and filtered through a pad ofCelite. The colorless filtrate was concentrated to dryness to give thetitle compound as a colorless oil. Yield: 430 mg (quantitative).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu), 4.78 (br s,2H, NH₂), 5.61 (br s, 2H, NH₂), 6.22 (d, J=5.3, 1H, H_(Py)), 7.19-7.22(m, 3H), 7.44 (d, J=8.7, 2H, H_(arom)), 9.43 (s, 1H, NHBoc); ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 28.1, 79.2, 115.3, 119.0, 122.8, 125.1,128.8, 131.7, 134.9, 139.1, 148.4, 152.5; LC-MS (m/z): 333.2 (M+H, 100),rt=3.06; HRMS (3.98 min): m/z calcd. for C₁₆H₂₁N₄O₂S [M+H⁺]: 333.13797.Found: 333.13812.

(IV). Cyclisation En-Route to Common Intermediates 1. Cyclisation toPyridopyrazin-3-One and Pyridopyrazine-2-One Synthesis 24 Tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Tert-butyl4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Method D1: tert-butyl 4-(2,3-diaminopyridin-4-yloxy)phenylcarbamate(0.86 g, 2.71 mmol) was dissolved in 15 ml of dry ethanol; 0.8 ml (4mmol) of a 50% ethyl glyoxalate solution in toluene were added and thesolution was stirred overnight at room temperature under Argonatmosphere. The solvent was partially evaporated, and tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (0.430g, 45% yield) is precipitated by addition of acetone (10 ml) andfiltered off.

tert-butyl4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (0.200g, 21% yield) is isolated by column chromatography over silica gel,eluant dichloromethane: ethyl acetate 1:1 Rf=0.3.

tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate: ¹H-NMR(DMSO-d₆), δ (ppm), J (Hz): 1.49 (s, 9H), 6.76 (d, 1H, J=5.4 Hz), 7.15(d, 2H, J=9.0 Hz), 7.57 (d, 2H, J=9.0 Hz), 8.32 (d, 1H, J=5.0 Hz), 8.40(s, 1H), 9.44 (bs, 1H), 12.54 (bs, 1H). LC-MS (m/z): 367 (M+H, 100).

tert-butyl4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate: ¹H-NMR(CDCl₃), δ (ppm), J (Hz): 1.54 (s, 9H), 6.55 (d, 1H, J=5.5 Hz), 6.67(bs, 1H), 7.14 (d, 2H, J=8.5 Hz), 7.49 (d, 2H, J=8.5 Hz), 8.36 (s, 1H),8.46 (d, 1H, J=5.5 Hz), 12.88 (bs, 1H). LC-MS (m/z): 367 (M+H, 100).

Synthesis 25 Tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate

Tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate

Method D1 was used with tert-butyl4-(2,3-diaminopyridin-4-yloxy)naphthalen-1-ylcarbamate (3.1 g) to affordthe title compounds Tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate(1.45 g, 42% yield) and Tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate(0.24 g, 9% yield).

Tert-butyl4-(2-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate: ¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.51 (s, 9H), 6.63 (d, 1H,5.6 Hz), 7.41 (d, 1H, J=8.3 Hz), 7.56 (m, 1H), 7.62 (m, 1H), 7.64 (d,1H, J=8.3 Hz), 7.90 (d, 1H, 7.7 Hz), 8.14 (d, 1H, 7.7 Hz), 8.25 (d, 1H,J=5.6 Hz), 8.45 (s, 1H), 9.39 (bs, 1H), 12.86 (bs, 1H). LC-MS (m/z): 405(M+H, 100).

Tert-butyl4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate: ¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.52 (s, 9H), 6.38 (d, 1H,5.7 Hz), 6.64 (d, 1H, J=8.2 Hz), 7.37 (d, 1H, J=6.6 Hz), 7.51-7.64 (m,2H), 7.83 (d, 1H, J=8.2 Hz), 8.14 (d, 1H, 6.6 Hz), 8.25 (s, 1H), 8.27(d, 1H, J=5.7 Hz), 9.38 (bs, 1H), 13.00 (bs, 1H). LC-MS: m/z: LC-MS(m/z): 405 (M+H, 60), 349 (100).

Synthesis 26Tert-butyl-2-(methylthio)-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Tert-butyl2-(methylthio)-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenylcarbamate

Using Method D1 with tert-butyl4-(2,3-diaminopyridin-4-yloxy)-2-(methylthio)phenyl carbamate (780 mg,2.15 mmol),tert-butyl-2-(methylthio)-4-(3-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (134 mg, 15% yield) andtert-butyl-2-(methylthio)-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (427 mg, 50% yield) were obtained.

Tert-butyl-2-(methylthio)-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate: ¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu);2.40 (s, 3H, CH₃); 6.87 (d, 1H, H_(Py) J=5.3 Hz), 7.01 (dd, 1H,H_(arom), J=8.6 Hz, J=2.6 Hz), 7.18 (d, 1H, H_(arom), J=2.6 Hz), 7.37(d, 1H, H_(arom), J=8.6 Hz), 8.36 (d, 1H, H_(Py) J=5.3 Hz), 8.42 (s, 1H,NH or CH), 8.46 (s, 1H, NH or CH), 12.57 (s, 1H, NH). ¹³C-NMR (DMSO-d₆),δ (ppm), J (Hz): 14.9, 27.9, 78.9, 110.1, 116.8, 118.0, 127.4, 132.6,137.0, 151.3, 153.4. LC-MS (m/z): 433 (M+H+MeOH, 100), rt=4.42 min.

Tert-butyl-2-(methylthio)-4-(3-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate: ¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.45 (s, 9H, tert-Bu);2.40 (s, 3H, CH₃); 6.59 (d, 1H, H_(Py) J=5.6 Hz), 6.98 (dd, 1H,H_(arom), J=8.6 Hz, J=2.6 Hz), 7.16 (d, 1H, H_(arom), J=2.6 Hz), 7.36(d, 1H, H_(arom), J=8.6 Hz), 8.17 (s, 1H, NH or CH), 8.36 (d, 1H,H_(Py), J=5.6 Hz), 8.44 (s, 1H, NH or CH), 12.89 (s, 1H, NH). ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 14.8, 27.9, 78.9, 106.2, 116.7, 117.8,118.2, 127.5, 132.5, 137.2, 145.4, 150.9, 151.5, 152.0, 153.4, 156.3,160.5. LC-MS (m/z): 401 (M+H, 100), rt=4.65 min.

Synthesis 27 Tert-butyl2-fluoro-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Tert-butyl2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenylcarbamate

Using Method D1 with tert-butyl4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenyl carbamate (3.50 g, 10.5mmol), tert-butyl2-fluoro-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (2.71 g, 69%) and tert-butyl2-fluoro-4-(3-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (0.96 g, 25%) were obtained.

Tert-butyl2-fluoro-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate:¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): δ=12.58 (br s, 1H, NHAr), 9.03 (br s,1H, NH_(Boc)), 8.41 (s, 1H, H_(arom)), 8.37 (d, J=5.5 Hz, 1H, H_(Py)),7.66 (vt, J=8.5 Hz, 1H, H_(arom)), 7.24 (d, 1H, H_(arom)), 7.06 (d, 1H,H_(arom)), 6.94 (d, J=5.5 Hz, 1H, H_(Py)), 1.47 ppm (s, 9H, tert-Bu);¹³C NMR (126 MHz, DMSO-d₆): δ=155.8 (br), 154.6, 154.5 (d, J_(FC)=248Hz), 153.1, 151.8 (br), 150.2 (d, J_(FC)=10 Hz), 145.4, 144.3 (br),125.7, 124.0 (d, J_(FC)=12 Hz), 119.9 (br), 116.1 (d, J_(FC)=3 Hz),110.7, 108.7 (d, J_(FC)=23 Hz), 79.4, 28.0 ppm; ¹⁹F NMR (470 MHz,DMSO-d₆): δ=−119.9 ppm; LC-MS (m/z): 373.4 (M+H, 100), rt=4.20 min; HRMS(5.15 min): m/z calcd. for C₁₈H₁₈FN₄O₄ [M+H⁺]: 373.13066. Found:373.13099.

Tert-butyl2-fluoro-4-(3-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate: ¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): δ=12.90 (br s, 1H, NHAr),9.01 (br s, 1H, NHBoc), 8.38 (d, J=5.5 Hz, 1H, H_(Py)), 8.17 (s, 1H,H_(arom)), 7.66 (vt, J=8.5 Hz, 1H, H_(arom)), 7.22 (d, 1H, H_(arom)),7.01 (d, 1H, H_(arom)), 6.67 (d, J=5.5 Hz, 1H, H_(Py)), 1.47 ppm (s, 9H,tert-Bu); ¹³C NMR (126 MHz, DMSO-d₆): δ=160.2, 156.4, 154.6 (d,J_(FC)=249 Hz), 153.1, 152.2, 151.2, 150.5 (d, J_(FC)=10 Hz), 145.6,125.8, 123.9 (d, J_(FC)=12 Hz), 118.5, 116.0 (d, J_(FC)=3 Hz), 108.5 (d,J_(FC)=23 Hz), 106.8, 79.4, 28.0 ppm; ¹⁹F NMR (470 MHz, DMSO-d₆):δ=−119.8 ppm; LC-MS (m/z): 373.1 (M+H, 100), rt=4.40 min; HRMS (5.34min): m/z calcd. for C₁₈H₁₇FN₄O₄ [M+H⁺]: 373.13066. Found: 373.13071.

Synthesis 28tert-butyl-3-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

tert-butyl-3-fluoro-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Using Method D1 with tert-butyl4-(2,3-diaminopyridin-4-yloxy)-3-fluorophenylcarbamate (1 g, 2.99 mmol),a mixture of two isomers (1.01 g, 90%) was obtained in a ratio 53/47.The crude powder was purified by Biotage to affordtert-butyl-3-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate(185 mg, 17% yield) andtert-butyl-3-fluoro-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate(370 mg, 34% yield) as white off powders.

tert-butyl-3-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate:¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.55 (s, 9H, tert-Bu); 6.56 (d, 1H,H_(Py) J=5.7 Hz), 7.37 (m, 2H, H_(arom)), 7.67 (m, 1H, H_(arom)), 8.22(s, 1H, CH), 8.37 (d, 1H, H_(Py), J=5.7 Hz), 9.75 (s, 1H, NH), 12.95 (s,1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.9, 80.6, 106.0, 107.4,115.6, 118.7, 124.6, 135.4, 139.8, 146.4, 152.3, 153.2, 153.7, 155.1,157.5, 161.5. ¹⁹F-NMR (δ, ppm, DMSO-d₆): −128.42. LC-MS (m/z): 373 (M+H,100), rt=2.43 min.

tertbutyl-3-fluoro-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.53 (s, 9H, tert-Bu); 6.83 (d, 1H,H_(Py) J=5.4 Hz), 7.32-7.41 (m, 2H, H_(arom)), 7.67 (m, 1H, H_(arom)),8.37 (d, 1H, H_(Py) J=5.4 Hz), 8.45 (s, 1H, CH), 9.75 (s, 1H, NH), 12.65(s, 1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.9, 80.6, 107.4,110.0, 115.6, 124.6, 135.4, 139.8, 146.4, 152.3, 153.0, 153.9, 155.4,157.5, 161.5. ¹⁹F-NMR (δ, ppm, DMSO-d₆): −128.12. LC-MS (m/z): 373 (M+H,100), rt=2.33 min.

Synthesis 29 Tert-butyl3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Tert-butyl3-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Using Method D1 with4-(3-N-(tert-butoxycarbonyl)aminophenoxy)-2,3-diamino-pyridine (1.00 g,3.16 mmol) tert-butyl3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (274mg, 24%) and tert-butyl3-(2-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (445mg, 1.26 mmol, 40%) were obtained.

tert-butyl3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate:

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu), 6.59 (d, 1H,H_(Py), J=5.6 Hz), 6.81-6.83 (m, 1H, H_(arom)), 7.36-7.39 (m, 3H,H_(arom)), 8.17 (s 1H, H_(arom)), 8.35 (d, 1H, H_(Py),6, J=5.6 Hz), 9.56(s, 1H, NHBoc), 12.89 (s, 1H, NH_(lactame)). LC-MS (m/z): 299 (M+H,100).

tert-butyl3-(2-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate:

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu), 6.84 (ddd, 1H,H_(arom), J=7.5 Hz, J=2.4 Hz, J=1.5 Hz), 6.86 (d, 1H, H_(Py), J=5.4 Hz),7.33-7.39 (m, 2H, H_(arom)), 7.42 (s, 1H, H_(arom)) 8.36 (d, 1H, H_(Py),J=5.4 Hz), 8.41 (s 1H, H_(arom)), 9.57 (s, 1H, NH_(Boc)), 12.54 (s, 1H,NH_(lactame)). LC-MS (m/z): 299 (M+H, 100).

Synthesis 30 Methyl3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)benzoate

Methyl 3-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)benzoate

Method D1 was used with methyl 3-(2,3-diaminopyridin-4-yloxy)benzoate(1.00 g, 3.86 mmol) to afford methyl3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)benzoate (402 mg, 35%)and methyl 3-(2-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)benzoate(750 mg, 2.52 mmol, 65%).

Methyl 3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)benzoate: ¹H-NMR(DMSO-d₆), δ (ppm), J (Hz): 3.85 (s, 3H, OMe), 6.68 (d, 1H, H_(Py),J=5.6 Hz), 7.53 (ddd, 1H, H_(arom), J=8.2 Hz, J=2.5 Hz, J=1.0 Hz), 7.65(t, 1H, H_(arom), J=8.0 Hz), 7.68 (dd, 1H, H_(arom), J=2.3 Hz, J=1.6 Hz)7.88 (ddd, 1H, H_(arom), J=7.7 Hz, J=2.5 Hz, J=1.2 Hz), 8.17 (s 1H,H_(arom)), 8.39 (d, 1H, H_(Py) J=5.6 Hz), 12.93 (s, 1H, NH). LC-MS(m/z): 298 (M+H, 100).

Methyl 3-(2-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)benzoate: ¹H-NMR(DMSO-d₆), δ (ppm), J (Hz): 3.86 (s, 3H, OMe), 6.97 (d, 1H, H_(Py),5,J=5.3 Hz), 7.56 (ddd, 1H, H_(arom), J=8.1 Hz, J=2.5 Hz, J=0.8 Hz), 7.66(t, 1H, H_(arom), J=8.0 Hz), 7.74 (dd, 1H, H_(arom), J=2.1 Hz, J=1.8 Hz)7.89 (d, 1H, H_(arom), J=7.8 Hz), 8.39 (d, 1H, H_(Py),6, J=5.3 Hz), 8.43(s 1H, H_(arom)), 12.58 (s, 1H, NH). LC-MS (m/z): 298 (M+H, 100).

Synthesis 31 Tert-butyl4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-ylthio)phenylcarbamate

Method D1 was used with Tert-butyl4-(2,3-diaminopyridin-4-ylthio)phenylcarbamate (1.058 g, 3.18 mmol) toafford the title compound as a yellow solid. Yield: 640 mg (54%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.50 (s, 9H, tert-Bu), 6.35 (d,J=5.4, 1H, H_(Py)), 7.52 (d, J=8.7, 2H, H_(arom)), 7.67 (d, J=8.7, 2H,H_(arom)), 8.19 (d, J=5.4, 1H, H_(Py)), 8.20 (s, 1H, H_(arom)), 9.70 (s,1H, NH_(Boc)), 12.84 (br s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz):28.1, 79.6, 114.6, 119.2, 119.5, 123.0, 136.7, 141.7, 143.3, 150.0,150.9, 152.5, 152.6, 156.7; LC-MS (m/z): 371.1 (m+H, 100), rt=4.97 min).

2. Cyclisation to Pyridopyrazin-2-Methyl-3-One andPyridopyrazine-3-Methyl-2-One Synthesis 32 Tert-butyl2-fluoro-4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Tert-butyl2-fluoro-4-(2-methyl-3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenylcarbamate

Method D2. Tert-butyl4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenylcarbamate (300 mg, 0.9mmol) was dissolved in dry EtOH (5 mL) and ethyl pyruvate (1 mL, 9 mmol)was added at once. After stirring for 16 h at RT, the precipitate wasfiltered and the two isomers were separated by column chromatography(silica gel, EtOAc as eluent).

Tert-butyl2-fluoro-4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate: 200 mg (58%). ¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.47 ppm (s,9H, tert-Bu); 2.48 (s, 3H, CH₃), 6.88 (d, 1H, J=5.5 Hz, H_(Py)), 7.03(d, 1H, H_(arom)), 7.22 (d, 1H, H_(arom)), 7.66 (vt, J=8.5 Hz, 1H,H_(arom)), 8.32 (d, J=5.3 Hz, H_(Py)), 9.00 (br s, 1H, NHBoc), 12.41 (brs, 1H, NHAr); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.0, 79.4, 106.8,109.0, 116.0, 118.5, 123.9, 125.8, 145.6, 150.5, 151.2, 152.2, 153.1,156.4, 160.3, ppm;

¹⁹F NMR (470 MHz, DMSO-d₆): δ=−119.9 ppm; LC-MS (m/z): 331.1.(M+H-tert-Bu, 100), rt=4.36 min.

Tert-butyl2-fluoro-4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate: 130 mg (38%). ¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.47 ppm (s,9H, tert-Bu), 2.42 (s, 3H, CH₃), 6.60 (d, 1H, J=5.5 Hz, H_(Py)), 7.03(d, 1H, H_(arom)), 7.22 (d, 1H, H_(arom)), 7.66 (vt, ³J_(FH)=8.5 Hz, 1H,H_(arom)), 8.30 (d, J=5.3 Hz, H_(Py)), 9.00 (br s, 1H, NH_(Boc)), 12.77(br s, 1H, NH_(arom)). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.0, 79.4,106.8, 109.0, 116.0, 118.5, 123.9, 125.8, 145.6, 150.5, 151.2, 152.2,153.1, 156.4, 160.3. ¹⁹F NMR (470 MHz, DMSO-d₆): δ=−119.9 ppm; LC-MS(m/z): m/z 331.1 (M+H-tert-Bu, 100), rt=4.55 min

Synthesis 33 tert-butyl4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenylcarbamate

Using Method D2 with tert-butyl4-(2,3-diaminopyridin-4-yloxy)-2-(methylthio) phenylcarbamate (570 mg,1.57 mmol), a mixture of two isomers was obtained. After cooling down,the crude was filtered, washed with ethanol and dried. The titlecompound (131 mg) was obtained as a white powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.45 (s, 9H, tert-Bu); 2.39 (s, 3H,CH₃); 2.43 (s, 3H, CH₃); 6.52 (d, 1H, H_(Py) J=5.6 Hz), 6.97 (dd, 1H,H_(arom), J=8.6 Hz, J=2.6 Hz), 7.15 (d, 1H, H_(arom), J=2.6 Hz), 7.36(d, 1H, H_(arom), J=8.6 Hz), 8.27 (d, 1H, H_(Py)J=5.6 Hz), 8.44 (s, 1H,NHBoc), 12.75 (s, 1H, NH). LC-MS (m/z): 415 (M+H, 100), rt=4.78 min.

Synthesis 34 tert-butyl4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenylcarbamate

Using Method D2 with tert-butyl4-(2,3-diaminopyridin-4-yloxy)-2-(methylthio) phenylcarbamate (570 mg,1.57 mmol), a mixture of the two isomers was obtained. The crude waspurified on silica gel (eluent: pure EtOAc), to afford the titlecompound (206 mg) as a pale yellow powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.45 (s, 9H, tert-Bu); 2.43 (s, 3H,CH₃); 2.48 (s, 3H, CH₃); 6.83 (d, 1H, H_(Py) J=5.2 Hz), 6.98 (dd, 1H,H_(arom), J=8.6 Hz, J=2.6 Hz), 7.15 (d, 1H, H_(arom), J=2.6 Hz), 7.35(d, 1H, H_(arom), J=8.6 Hz), 8.31 (d, 1H, H_(Py) J=5.2 Hz), 8.45 (s, 1H,NH), 12.51 (bs, 1H, NH). LC-MS (m/z): 531 (M+H+C₅H₈O₃, 100), rt=4.78min.

Synthesis 35 tert-butyl3-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

tert-butyl3-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Method D2 was used with4-(3-N-(tert-butoxycarbonyl)aminophenoxy)-2,3-diamino-pyridine to afforda mixture of the 2 isomers. The mixture was chromatographied (eluent:CH₂Cl₂/EtOAc: 1/0 towards 0/1) to afford at first the tert-butyl3-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate as a yellow solid (194 mg, 0.527 mmol, 11%) and thentert-butyl3-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate as a yellow solid (841 mg, 2.28 mmol, 48%).

tert-butyl3-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate:

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu), 2.43 (s, 3H,Me), 6.53 (d, 1H, H_(Py),5, J=5.6 Hz), 6.81-6.83 (m, 1H, H_(arom)),7.36-7.37 (m, 3H, H_(arom)), 8.27 (d, 1H, H_(Py),6, J=5.6 Hz), 9.56 (s,1H, NH_(Boc)), 12.75 (s, 1H, NH_(lactame)). LC-MS (m/z): 369 (M+H, 100).

tert-butyl3-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate:

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.46 (s, 9H, tert-Bu), 2.48 (s, 3H,Me), 6.80-6.83 (m, 2H, H_(arom)), 7.32-7.37 (m, 2H, H_(arom)), 7.40 (s,1H, H_(arom)), 8.31 (d, 1H, H_(Py), J=5.4 Hz), 9.55 (s, 1H, NH_(Boc)),12.38 (s, 1H, NH_(lactame)). LC-MS (m/z): 369 (M+H, 100).

Synthesis 36 tert-butyl4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate

tert-butyl 4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate

Method D2 was used with tert-butyl4-(2,3-diaminopyridin-4-yloxy)naphthalen-1-ylcarbamate to afford amixture of isomers. The residue was chromatographied (eluent:CH₂Cl₂/EtOAc: 6/1 towards 0/1 then EtOAc/MeOH: 95/5) to afford at firsttert-butyl 4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate as a slightly yellow solid (401 mg, 0.958 mmol,35%) and then tert-butyl4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate as a yellow solid (607 mg, 1.45 mmol, 53%).

tert-butyl 4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate: ¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.53 (s,9H, tert-Bu), 2.01 (s, 3H, Me), 6.32 (d, 1H, H_(Py) J=5.7 Hz), 7.38 (d,1H, H_(arom), J=8.2 Hz), 7.55-7.58 (m, 1H, H_(arom)), 7.62-7.67 (m, 2H,H_(arom)), 7.85 (d, 1H, H_(arom), J=8.4 Hz), 8.17 (d, 1H, H_(arom),J=8.6 Hz), 8.20 (d, 1H, H_(Py), J=5.6 Hz), 9.35 (s, 1H, NH_(Boc)), 12.82(s, 1H, NH_(lactame)). ¹³C-NMR (5, ppm, DMSO-d₆): 20.50 (CH₃), 28.05(tert-Bu), 79.03 (tert-Bu), 105.56, 116.89, 117.37, 121.10, 121.13,123.56, 126.26, 126.52, 126.79, 129.23, 132.08, 145.63, 146.03, 150.51,153.98, 156.28, 159.14, 160.49. LC-MS (m/z): 419 (M+H, 100).

tert-butyl 4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate: ¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.51 (s,9H, tert-Bu), 2.52 (s, 3H, Me), 6.58 (d, 1H, H_(Py),5, J=5.4 Hz), 7.37(d, 1H, H_(arom), J=8.2 Hz), 7.53-7.64 (m, 2H, H_(arom)), 7.91 (d, 1H,H_(arom), J=8.1 Hz), 8.14 (d, 1H, H_(arom), J=8.5 Hz), 8.22 (d, 1H,H_(Py),6, J=5.4 Hz), 9.32 (s, 1H, NH_(Boc)), 12.66 (s, 1H,NH_(lactame))_(.) ¹³C-NMR (δ, ppm, DMSO-d₆): 20.93 (CH₃), 28.05(tert-Bu), 79.01 (tert-Bu), 108.55, 116.54, 118.89, 121.02, 121.48,123.38, 126.23, 126.53, 126.59, 129.24, 132.03, 143.82, 144.89, 145.87,152.08, 153.97, 154.52, 164.03. LC-MS (m/z): 419 (M+H, 100).

3. Cyclisation to Pyridopyrazin-2,3-Dione Synthesis 378-(4-aminophenoxy)pyrido[2,3-b]pyrazine-2,3(1H,4H)-dione

Method D3. A solution of tert-butyl 4-(2,3-diaminopyridin-4-yloxy)phenylcarbamate (0.320 g, 1.0 mmol) in diethyl oxalate(2 ml) is reacted twice for 10 minutes in a microwave reactor (180 C,150 W). The solution is cooled and the solid filtered and washed withcold ethanol. Obtained8-(4-aminophenoxy)pyrido[2,3-b]pyrazine-2,3(1H,4H)-dione (70 mg, 25%yield) as a grey solid.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.36 (d, 1H, J=5.7 Hz), 6.67 (d, 2H,J=8.6 Hz), 6.88 (d, 2H, J=8.6 Hz), 7.82 (d, 2H, J=5.7 Hz), 11.76 (bs,1H), 12.28 (bs, 1H). LC-MS (m/z): 271 (M+H, 100).

Synthesis 388-(4-Amino-3-fluorophenoxy)pyrido[3,2-b]pyrazine-2,3(1H,4H)-dione

A solution of tert-butyl4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenylcarbamate (1.03 g, 3.08mmol) was dissolved in dry EtOH (10 mL), diethyl oxalate (10 mL) wasadded and the solution was heated to reflux for 96 h, cooled to RT andfiltered. The title compound was isolated as a white solid. Yield: 820mg (92%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.20 (br s, 2H, NH₂), 6.44 (d, J=5.7,1H, H_(Py)), 6.79 (m, 1H, H_(arom)), 6.85 (m, 1H, H_(arom)), 6.98 (m,1H, H_(arom)), 7.91 (d, J=5.7, 1H, rH_(Py)), 11.81 (s, 1H, NH), 12.34(s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 107.4, 108.4 (d,J_(FC)=21.9), 113.0, 116.2 (d, J_(FC)=3.0), 121.9 (br), 123.0 (br),140.7, 143.2, 149.6 (d, J_(FC)=10.1), 151.2, 153.8 (d, J_(FC)=240),154.8, 155.9; ¹⁹F-NMR (DMSO-d₆), δ (ppm), J (Hz): −123.5 ppm; LC-MS(m/z): 289.1 (M+H, 100).

4. Cyclisation to 2-Aminopyridopyrazin-3-One Synthesis 39 tert-butyl4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate

tert-butyl4-(3-amino-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate

Method D4. To tert-butyl4-(2,3-diaminopyridin-4-yloxy)naphthalen-1-ylcarbamate (1.16 g, 3.17mmol) dissolved in 15 mL of anhydrous ethanol under argon was added theethyl carboethoxyformimidate hydrochloride (1.72 g, 9.51 mmol). Thereaction mixture was stirred at reflux for 48 hours. After cooling atRT, a precipitate was formed. It was collected and rinsed with ether.The first isomer was obtained as a slightly pink solid (275 mg, 21%).Solvent was evaporated under vacuum and the residue was retaken inEtOAc. The organic phases were washed with a saturated solution ofNaHCO₃, then brine, dried over MgSO₄ and concentrated under vacuum. Theresidue was chromatographied (eluent: EtOAc/MeOH: 1/0 towards 9/1) toafford the second isomer as a slightly yellow solid (463 mg, 35%).

tert-butyl4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate:¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.51 (s, 9H, tert-Bu), 6.31 (d, 1H,H_(Py),5, J=5.6 Hz), 7.20 (d, 1H, H_(arom), J=8.2 Hz), 7.52-7.62 (m, 3H,H_(arom)), 7.89-7.91 (m, 2H, H_(arom)), 8.10 (d, 1H, H_(arom), J=8.4Hz), 9.26 (s, 1H, NHBoc), 12.61 (s, 1H, NH_(lactame)). ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 28.06 (C(CH₃)), 78.91 (C(CH₃)), 106.62,115.63, 119.47, 121.28, 121.58, 123.43, 126.26, 126.42, 126.48, 129.44,131.17, 142.99, 143.57, 147.15, 151.74, 152.72, 154.07, 156.97. LC-MS(m/z): 420 (M+H, 100).

Second isomer tert-butyl4-(3-amino-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate:¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.51 (s, 9H, tert-Bu), 6.28 (d, 1H,H_(Py), J=5.5 Hz), 7.30 (d, 1H, H_(arom), J=8.2 Hz), 7.62-7.53 (m, 2H,H_(arom)), 7.95 (d, 1H, H_(arom), J=8.4 Hz), 8.00 (d, 1H, H_(arom),J=5.5 Hz), 8.04 (t, 1H, H_(arom), J=8.3 Hz), 8.12 (d, 1H, H_(arom),J=8.7 Hz), 9.29 (s, 1H, NHBoc), 12.41 (s, 1H, NH_(lactame)). ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 28.17 (tert-Bu), 79.06 (tert-Bu), 104.81,114.12, 116.17, 119.24, 121.27, 121.77, 123.40, 126.52, 126.57, 129.39,131.69, 144.40, 146.51, 146.90, 151.12, 151.13, 154.13, 154.89.

5. Conversion of Pyridopyrazin-2-One and Pyridopyrazin-3-One to2-Amino-Pyridopyrazine and 3-Amino-Pyridopyrazine Synthesis 40Tert-butyl4-(3-chloropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenylcarbamate

Method D5: N-chloro succinimide (91 mg, 681 μmol) was added to asolution of triphenyl phosphine (178 mg, 678 μmol) in dry 1,4-dioxane (4mL) under Ar, yielding a white suspension. After 30 min, tert-butyl2-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate(48 mg, 129 μmol) was added at once and the mixture was heated to refluxfor 1 h. The black mixture was cooled to RT, Et₃N (1 mL) was added, andall volatiles were evaporated. The black residue was dissolved in CH₂Cl₂(3 mL) and loaded onto a silica gel column (packed with Et₂O). Elutionwith ether furnished the title compound as the first, fast-running band(R_(f)=0.83 in Et₂O), which was concentrated to dryness to a whitesolid. Yield: 34 mg (68%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.48 (s, 9H, tert-Bu), 7.09-7.13 (m,2H), 7.32 (m, 1H, H_(arom)), 7.71 (m, 1H, H_(arom)), 8.98 (d, 1H, J=5.3,H_(Py)), 9.06 (s, 1H, NH_(Boc)), 9.12 (s, 1H, H_(arom)); ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 28.0, 79.5, 108.8 (d, J_(FC)=23.1), 109.9,116.2 (d, J_(FC)=3.1), 124.3 (d, J_(FC)=11.6), 125.8, 129.3, 145.3,149.8, 150.2 (d, J_(FC)=10.3), 150.8, 153.1, 154.6 (d, J_(FC)=248),156.1, 161.0; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −119.6; LC-MS (m/z): 391.1(M+H, 100), rt=4.40 min.

Synthesis 41 Tert-butyl4-(2-chloropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenylcarbamate

Method D5 was used with tert-butyl2-fluoro-4-(2-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamateto give the title product as off-white crystals. Yield: 250 mg (50%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.48 (s, 9H, tert-Bu), 7.09-7.14 (m,2H, H_(arom)), 7.34 (m, 1H, H_(arom)), 7.73 (m, 1H, H_(arom)), 8.97 (d,1H, J=5.3, _(Py)rH), 9.07 (s, 1H, NHBoc), 9.23 (s, 1H, H_(arom))_(;)¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.0, 79.5, 109.0 (d, J_(FC)=23.1),110.0, 116.4 (d, J_(FC)=3.1), 124.5 (d, J_(FC)=11.6), 125.7, 129.8,146.6, 149.0, 149.8 (d, J_(FC)=10.3), 150.7, 153.1, 154.6 (d,J_(FC)=248), 155.0, 160.1; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −119.6; LC-MS(m/z): 391.1 (M+H, 100), rt=4.80 min.

Synthesis 42 Tert-butyl2-fluoro-4-(2-morpholinopyrido[2,3-b]pyrazin-8-yloxy)phenyl-carbamate

Method D6: Morpholine (500 μL, excess) was added to tert-butyl4-(2-chloropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenylcarbamate (68 mg,174 μmol) under argon and the yellow solution was stirred at RT for 45min. Next, H₂O (10 mL) was added and the precipitated yellow solid wasfiltered to give the title compound as a yellow solid. Yield: 69 mg(90%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.47 (s, 9H, tert-Bu), 3.69 (br s,8H, N(CH₂CH₂)₂O) 6.96 (m, 2H, H_(arom)), 7.04 (d, 1H, J=5.3, H_(Py)),7.15 (m, 1H, H_(arom)), 7.59 (m, 1H, H_(arom)), 8.57 (d, 1H, J=5.3,H_(Py)), 8.96 (s, 1H, NH_(Boc)), 8.98 (s, 1H, H_(arom))_(;) ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 28.0, 44.5, 65.8, 79.3, 107.8 (d,J_(FC)=23.1), 111.5, 115.3 (d, J_(FC)=3.1), 122.9 (d, J_(FC)=11.6),125.8, 128.8, 139.5, 147.8, 147.9, 151.1, 152.3 (d, J_(FC)=10.3), 153.2,154.8 (d, J_(FC)=248), 157.6; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −120.3; LC-MS(m/z): 442.2 (M+H, 100) rt=4.70 min.

Synthesis 43 Tert-butyl2-fluoro-4-(3-morpholinopyrido[2,3-b]pyrazin-8-yloxy)phenyl-carbamate

Method D6 was used with morpholine and tert-butyl4-(3-chloropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenylcarbamate to givethe title compound as a white solid. Yield: 117 mg (89%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.48 (s, 9H, tert-Bu), 3.77 (m, 4H,N(CH₂CH₂)₂O), 3.84 (m, 4H, N(CH₂CH₂)₂O), 6.72 (d, 1H, J=5.3, H_(Py)),7.00 (m, 2H, H_(arom)), 7.21 (m, 1H, H_(arom)), 7.63 (m, 1H, H_(arom)),8.65 (d, 1H, J=5.3, H_(Py)), 8.85 (s, 1H, H_(arom)), 9.00 (s, 1H,NH_(Boc)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.0, 44.4, 65.9, 79.4,106.4, 108.2 (d, J_(FC)=23.1), 115.7 (d, J_(FC)=3.1), 122.6, 123.3 (d,J_(FC)=11.6), 125.9, 136.4, 146.7, 151.4 (d, J_(FC)=10.3), 152.2, 153.2,153.8, 153.9, 154.7 (d, J_(FC)=248), 160.0; ¹⁹F-NMR (DMSO-d₆), δ (ppm):−120.0; LC-MS (m/z): 442.2 (M+H, 100), rt=3.48 min.

Synthesis 44 Tert-butyl2-fluoro-4-(3-(methylamino)pyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate

Method D6 was used with methylamine and Tert-butyl4-(3-chloropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenylcarbamate to givethe title compound as a white solid. Yield: 80 mg (90%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.47 (s, 9H, tert-Bu), 2.95 (d,J=4.7, 3H, NHCH₃), 6.62 (d, J=5.4, 1H, H_(Py)), 6.97 (m, 1H, H_(arom)),7.18 (m, 1H, H_(arom)), 7.61 (m, 1H, H_(arom)), 8.03 (br q, J=4.7, 1 H,NHCH₃), 8.30 (s, 1H, H_(arom)), 8.55 (d, J=5.4, 1H, H_(Py)), 8.97 (s,1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 27.1, 28.0, 79.4, 105.6,108.3 (d, J_(FC)=23.0), 115.8 (d, J_(FC)=2.9), 122.3, 123.4 (d,J_(FC)=11.9), 125.8 (br), 139.7 (br), 151.3 (d, J_(FC)=10.0), 152.8,153.2, 153.4, 154.7 (d, J_(FC)=248), 155.3, 160.1; ¹⁹F-NMR (DMSO-d₆), δ(ppm): −120.0; LC-MS (m/z): 386.1 (M+H, 100), rt=3.13 min.

Synthesis 45 Tert-butyl2-fluoro-4-(3-(4-methylpiperazin-1-yl)pyrido[3,2-b]pyrazin-8-yloxy)phenylcarbamate

Method D6 was used with N-methylpiperazine and tert-butyl4-(3-chloropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenylcarbamate to givethe title compound as a yellow solid. Yield: 142 mg (92%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.47 (s, 9H, tert-Bu), 2.24 (s, 3H,CH₃), 2.46 (m, 4H, N(CH₂CH₂)₂NMe), 3.84 (m, 4H, N(CH₂CH₂)₂NMe), 6.68 (d,J=5.3, 1H, H_(Py)), 6.98 (m, 1H, H_(arom)), 7.19 (m, 1H, H_(arom)), 7.61(m, 1H, H_(arom)), 8.62 (d, J=5.3, 1H, H_(Py)), 8.84 (s, 1H, H_(arom)),8.97 (s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.0, 43.9, 45.6,54.2, 79.4, 106.2, 108.2 (d, J_(FC)=22.8), 115.7 (d, J_(FC)=3.1), 122.4,123.4 (d, J_(FC)=11.9), 125.9, 136.4, 151.4 (d, J_(FC)=10.0), 152.3,153.2, 153.7, 154.7 (d, J_(FC)=248), 160.1;

¹⁹F-NMR (DMSO-d₆), δ (ppm): −120.0; LC-MS (m/z): 455.2 (M+H, 100),rt=2.43 min.

Synthesis 46 Tert-butyl4-(3-(dimethylamino)pyrido[3,2-b]pyrazin-8-yloxy)-2-fluorophenylcarbamate

Method D6 was used with tert-butyl4-(3-chloropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenylcarbamate (270mg, 0.67 mmol) and dimethylamine to give the product as a yellow solid.Yield: 233 mg (91%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.47 (s, 9H, tert-Bu), 2.95 (d,J=4.7, 3H, NHCH₃), 6.62 (d, J=5.4, 1H, H_(Py)), 6.97 (m, 1H, H_(arom)),7.18 (m, 1H, H_(arom)), 7.61 (m, 1H, H_(arom)), 8.03 (br q, J=4.7, 1 H,NHCH₃), 8.30 (s, 1H, H_(arom)), 8.55 (d, J=5.4, 1H, H_(Py)), 8.97 (s,1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.0, 37.4, 79.4, 105.8,108.2 (d, J_(FC)=23.0), 115.7 (d, J_(FC)=2.9), 122.0, 123.4 (d,J_(FC)=11.9), 125.9 (br), 136.1, 151.5 (d, J_(FC)=10.0), 152.5, 153.2,153.5, 154.7 (d, J_(FC)=248), 160.1; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −120.0;LC-MS (m/z): 400.1 (M+H, 100), rt=1.97 min.

6. Cyclisation to Other Substituted Pyridopyrazinones Synthesis 47tert-butyl4-(3-oxo-2-(trifluoromethyl)-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate

tert-butyl4-(2-oxo-3-(trifluoromethyl)-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate

Method D7. To tert-butyl4-(2,3-diaminopyridin-4-yloxy)naphthalen-1-ylcarbamate (1.00 g, 2.73mmol) dissolved in 20 mL of anhydrous ethanol under argon and at refluxwas added the ethyl trifluoropyruvate (697 mg, 0.50 mL, 4.10 mmol). Thereaction mixture was stirred at reflux for 3 hours. After cooling at RT,a precipitate was formed, filtered off and rinsed with Et₂O. tert-butyl4-(3-oxo-2-(trifluoromethyl)-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamatewas obtained as a white solid (116 mg, 0.246 mmol, 9%). The filtrate wasevaporated under vacuum. The residue was chromatographied (eluent:CH₂Cl₂/EtOAc: 4/1 towards 0/1) to afford the second isomer as a slightlyyellow solid (540 mg, 1.14 mmol, 42%).

First isomer tert-butyl4-(3-oxo-2-(trifluoromethyl)-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate:¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.52 (s, 9H, tert-Bu), 6.37 (d, 1H,H_(Py) J=5.7 Hz), 7.45 (d, 1H, H_(arom), J=8.2 Hz), 7.54-7.57 (m, 1H,H_(arom)), 7.62-7.65 (m, 1H, H_(arom)), 7.69 (d, 1H, H_(arom), J=8.2Hz), 7.80 (d, 1H, H_(arom), J=8.4 Hz), 8.18 (d, 1H, H_(arom), J=8.6 Hz),8.37 (d, 1H, H_(Py), J=5.7 Hz), 9.38 (s, 1H, NHBoc), 13.55 (s, 1H,NH_(lactame)). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 28.04 (tert-Bu),79.09 (tert-Bu), 105.71, 116.54, 117.35, 118.78, 120.84, 120.97, 123.62,126.06, 126.58, 126.97, 129.09, 132.63, 143.17, 145.24, 146.71, 153.20,153.90, 154.84, 162.26. LC-MS (m/z): 473 (M+H, 100).

Second isomer tert-butyl4-(2-oxo-3-(trifluoromethyl)-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamate:¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.53 (s, 9H, tert-Bu), 6.76 (d, 1H,H_(Py), J=5.3 Hz), 7.43 (d, 1H, H_(arom), J=8.2 Hz), 7.55-7.58 (m, 1H,H_(arom)), 7.63-7.69 (m, 2H, H_(arom)), 7.92 (d, 1H, H_(arom), J=8.4Hz), 8.17 (d, 1H, H_(arom), J=8.6 Hz), 8.39 (d, 1H, H_(Py) J=5.3 Hz),9.38 (s, 1H, NH_(Boc)), 13.51 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d₆),δ (ppm), J (Hz): 28.06 (tert-Bu), 79.08 (tert-Bu), 90.66 (CF₃), 110.72,116.81, 118.62, 120.88, 120.97, 121.42, 123.17, 123.46, 125.47, 126.07,126.68, 129.25, 132.38, 145.51, 146.71, 151.66, 153.97, 166.39. LC-MS(m/z): 473 (M+H, 100).

(V) Deprotection of Boc Synthesis 488-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-2(1H)-one

Method E1: Tert-butyl2-fluoro-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (250 mg, 671 μmol) was added to a round bottom flask under Ar.TBAF (7 mL of a 1M solution in THE, 7 mmol) was added and the solutionwas heated to reflux for 5 h. The volatiles were evaporated and the oilyresidue was diluted with H₂O (80 mL). The pH was adjusted to 7 (NaHCO₃)and after 1 h of stirring at RT, the precipitate was filtered off andstripped twice with toluene (30 mL) to give the title compound as ayellow solid. Yield: 180 mg (98%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.19 ppm (br s, 2H, NH₂), 6.79 (d,J=5.4 Hz, 1H, H_(Py)), 6.88-6.82 (m, 2H, H_(arom)), 7.06 (m, 1H,H_(arom)), 8.32 (d, J=5.4 Hz, 1H, H_(Py)), 8.40 (s, 1H, H_(arom)), 12.49(br s, 1H, NHAr). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 108. (d, J_(FC)=21Hz), 109.3, 116.3 (d, J_(FC)=6 Hz), 117.1 (d, J_(FC)=3 Hz), 119.2 (br),134.7 (d, J_(FC)=13 Hz), 142.6 (d, J_(FC)=9 Hz), 144.0 (br), 145.4,150.1 (d, J_(FC)=240 Hz), 153.2, 154.5, 155.6 (br); LC-MS (m/z): 273.1(M+H, 100), rt=2.37 min

Synthesis 49 8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one

Method E1 was used with tert-butyl2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenylcarbamate to afford the title compound. Yield: 191 mg (93%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.21 ppm (br s, 2H, NH₂); 6.52 (d,J=4.8 Hz, 1H, H_(Py)), 6.89-6.82 (m, 2H, H_(arom)), 7.05 (d,³J_(FC)=11.5 Hz, 1H, H_(arom)), 8.17 (s, 1H, H_(arom)), 8.32 (d, J=4.8Hz, 1H, H_(Py)), 12.86 (br s, 1H, NHAr); ¹³C-NMR (DMSO-d₆), δ (ppm), J(Hz): 105.5, 108.8 (d, J_(FC)=21 Hz), 116.4 (d, J_(FC)=6 Hz), 117.0 (d,J_(FC)=3 Hz), 118.0, 134.6 (d, J_(FC)=13 Hz), 142.6 (d, J_(FC)=9 Hz),145.3, 150.8, 150.1 (d, J_(FC)=241 Hz), 152.1, 156.5, 161.7; ¹⁹F NMR(470 MHz, DMSO-d₆): δ=−131.2 ppm; LC-MS (m/z): 273.1 (M+H, 100), rt=2.86min.

Synthesis 508-(4-amino-3-(methylthio)phenoxy)pyrido[2,3-b]pyrazin-2(1H)-one

Using Method E1 withtert-butyl-2-(methylthio)-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate(170 mg, 0.4 mmol), the title compound (81 mg, 63%) was obtained as apale brown powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.36 (s, 3H, CH₃); 5.19 (s, 2H, NH₂),6.75 (d, 1H, H_(Py) J=5.3 Hz), 6.79 (d, 1H, H_(arom), J=8.6 Hz), 6.90(dd, 1H, H_(arom), J=8.6 Hz, J=2.5 Hz), 7.07 (d, 1H, H_(arom), J=2.5Hz), 8.31 (d, 1H, H_(Py) J=5.3 Hz), 8.39 (s, 1H, NH or CH), 12.48 (s,1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 15.7, 108.9, 114.8, 119.8,120.7, 121.8, 143.6, 145.0, 145.3, 154.4. LC-MS (m/z): 301 (M+H, 100),rt=2.90 min.

Synthesis 518-(4-amino-3-(methylthio)phenoxy)pyrido[3,2-b]pyrazin-3(4H)-one

Using Method E1 with tert-butyl2-(methylthio)-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenylcarbamate(110 mg, 0.3 mmol), the title compound (63 mg, 76%) was obtained as apale yellow powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.37 (s, 3H, CH₃); 5.18 (5, 2H, NH₂),6.48 (d, 1H, H_(Py) J=5.6 Hz), 6.79 (d, 1H, H_(arom), J=8.6 Hz), 6.87(dd, 1H, H_(arom), J=8.6 Hz, J=2.6 Hz), 7.04 (d, 1H, H_(arom), J=2.6Hz), 8.16 (5, 1H, NH or CH), 8.29 (d, 1H, H_(Py) J=5.6 Hz), 12.82 (5,1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 15.6, 105.2, 114.8, 117.8,119.7, 120.8, 121.5, 143.7, 144.9, 145.1, 150.5, 151.9, 156.4, 161.8.LC-MS (m/z): 301 (M+H, 100), rt=3.35 min.

Synthesis 528-(4-amino-3-fluorophenoxy)-3-methylpyrido[3,2-b]pyrazin-2(1H)-one

Method E1 was used with tert-butyl2-fluoro-4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate; yield: 96%.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.48 ppm (s, 3H, CH₃), 6.72 (d, 1H,J=5.3 Hz, H_(Py)), 6.84 (m, 2H, H_(arom)), 7.03 (m, 1H, H_(arom)), 8.27(d, J=5.3 Hz, 1H, H_(Py)), 12.32 (br s, 1H, NHAr); ¹³C-NMR (DMSO-d₆), δ(ppm), J (Hz): 28.0, 79.4, 106.8, 109.0, 116.0, 118.5, 123.9, 125.8,145.6, 150.5, 151.2, 152.2, 153.1, 156.4, 160.3; ¹⁹F NMR (470 MHz,DMSO-d₆): δ=−131.3 ppm; LC-MS (2.79 min): 287.1 (M+H, 100)

Synthesis 538-(4-amino-3-fluorophenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one

Method E1 was used with tert-butyl2-fluoro-4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate to afford the title compound in 97% yield.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.43 ppm (s, 3H, CH₃), 5.18 (br s,2H, NH₂), 6.46 (d, J=4.8 Hz, 1H, H_(Py)), 6.81 (m, 1H, H_(arom)), 7.02(s, 1H, H_(arom)), 8.23 (d, J=4.8 Hz, 1H, H_(Py)), 12.70 (br s, 1H,NHAr); ¹⁹F NMR (470 MHz, DMSO-d₆): δ=−131.2 ppm; LC-MS (m/z): 287.1(M+H, 100), rt=3.20 min.

Synthesis 54 8-(4-amino-2-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one

Using Method E1 withtert-butyl-3-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate(335 mg, 0.9 mmol), the title compound (164 mg, 67%) was obtained as abrown powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.66 (bs, 2H, NH2); 6.57 (d, 1H,H_(Py), J=5.4 Hz), 6.72 (dd, 1H, H_(arom), J=8.7 Hz and J=2.0 Hz), 6.84(dd, 1H, H_(arom), J=12.6 Hz and J=2.5 Hz), 7.20 (t, 1H, H_(arom), J=8.8Hz), 8.21 (s, 1H, CH), 8.38 (d, 1H, H_(Py), J=5.7 Hz). ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 105.1, 106.0, 114.0, 118.7, 124.8, 133.0,145.0, 146.4, 152.1, 153.2, 155.8, 157.5, 161.9. ¹⁹F-NMR (δ, ppm,DMSO-d6): −129.18. LC-MS (m/z): 273 (M+H, 100), rt=1.45 min.

Synthesis 55 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one

Method E1 was used with tert-butyl3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (226mg, 0.638 mmol) to afford the title compound as a slightly yellow solid(132 mg, 0.519 mmol, 81%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.37 (bs, 2H, NH₂), 6.30 (ddd, 1H,H_(arom), J=7.9 Hz, J=2.3 Hz, J=0.7 Hz), 6.35 (t, 1H, H_(arom), J=2.2Hz), 6.50 (ddd, 1H, H_(arom), J=8.1 Hz, J=2.0 Hz, J=0.8 Hz), 6.58 (d,1H, H_(Py), J=5.6 Hz), 7.11 (t, 1H, H_(arom), J=8.0 Hz), 8.16 (s, 1H,H_(arom)), 8.33 (d, 1H, H_(Py) J=5.6 Hz), 12.84 (s, 1H, NH). ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 105.03, 106.21, 106.83, 111.13, 118.24,130.33, 145.30, 150.75, 150.78, 151.89, 154.70, 156.36, 160.82. LC-MS(m/z): 255 (M+H, 100).

Synthesis 56 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-2(1H)-one

Method E1 was used with tert-butyl3-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (635mg, 1.8 mmol) to afford the title compound as a slightly yellow solid(123 mg, 0.484 mmol, 27%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.38 (bs, 2H, NH₂), 6.37 (ddd, 1H,H_(arom), J=7.9 Hz, J=2.3 Hz, J=0.7 Hz), 6.35 (t, 1H, H_(arom), J=2.2Hz), 6.50 (ddd, 1H, H_(arom), J=8.1 Hz, J=2.0 Hz, J=0.8 Hz), 6.85 (d,1H, H_(Py), J=5.6 Hz), 7.11 (t, 1H, H_(arom), J=8.0 Hz), 8.16 (s, 1H,H_(arom)), 8.35 (d, 1H, H_(Py), J=5.3 Hz), 8.40 (5, 1H, H_(arom)), 12.49(5, 1H, NH). LC-MS (m/z): 255 (M+H, 100).

Synthesis 57 8-(3-aminophenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one

Method E1 was used with tert-butyl3-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate(190 mg, 0.5 mmol) to afford the title compound as a slightly yellowsolid (120 mg, 0.447 mmol, 90%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.42 (s, 3H, Me), 5.37 (bs, 2H, NH₂),6.31 (d, 1H, H_(arom), J=7.9 Hz), 6.36 (s, 1H, H_(arom)), 6.50 (d, 1H,H_(arom), J=8.0 Hz), 6.53 (d, 1H, H_(Py) J=5.7 Hz), 7.12 (t, 1H,H_(arom), J=8.0 Hz), 8.26 (d, 1H, H_(Py) J=5.7 Hz), 12.77 (5, 1H, NH).¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 20.49 (Me), 105.26, 105.96, 107.06,111.14, 117.72, 130.40, 145.96, 150.36, 150.84, 154.87, 156.61, 158.70,160.10. LC-MS (m/z): 269 (M+H, 100).

Synthesis 58 8-(4-aminophenylthio)pyrido[2,3-b]pyrazin-3(4H)-one

Method E1 was used with Tert-butyl4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-ylthio)phenylcarbamate (438mg, 1.18 mmol) to give the title compound as a yellow solid. Yield: 160mg (50%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.68 (br s, 2H, NH₂), 6.39 (d, J=5.3,1H, H_(Py)), 6.71 (d, J=8.3, 2H, H_(arom)), 7.22 (d, J=8.3, 2H,H_(arom)), 8.17 (m, 2H, H_(Py)), 12.78 (br s, 1H, NH); ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 110.0, 114.5, 115.2, 123.0, 137.1, 143.2,149.9, 150.7, 151.0, 154.1, 156.7; LC-MS (m/z): 271.0 (M+H, 100),rt=3.46 min.

Synthesis 592-fluoro-4-(3-morpholinopyrido[2,3-b]pyrazin-8-yloxy)aniline

Method E1 was used with Tert-butyl2-fluoro-4-(3-morpholinopyrido[2,3-b]pyrazin-8-yloxy)phenyl-carbamate(100 mg, 0.23 mmol) to give the title compound as a yellow solid. Yield:69 mg (87%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 3.76 (m, 4H, N(CH₂CH₂)₂O), 3.82 (m,4H, N(CH₂CH₂)₂O), 5.16 (s, 2H, NH₂), 6.52 (d, 1H, J=5.3, H_(Py)),6.80-6.88 (m, 2H, H_(arom)), 7.03 (m, 1H, H_(arom)), 8.55 (d, 1H, J=5.3,H_(Py)), 8.82 (s, 1H, H_(arom)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz):44.4, 65.9, 104.5, 108.7 (d, J_(FC)=21.2), 116.4 (d, J_(FC)=5.8), 117.0(d, J_(FC)=2.9), 122.3, 134.4 (d, J_(FC)=12.9), 135.9, 143.2 (d,J_(FC)=9.5), 150.2 (d, J_(FC)=240), 151.9, 153.2, 153.9, 161.6; ¹⁹F-NMR(DMSO-d₆), δ (ppm): −131.3; LC-MS (m/z): 342.1 (M+H, 100), rt=2.03 min.

Synthesis 608-(4-amino-3-fluorophenoxy)-N-methylpyrido[3,2-b]pyrazin-3-amine

Method E1 was used with tert-butyl2-fluoro-4-(3-(methylamino)pyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate(65 mg, 0.17 mmol) to give 41 mg of the title compound (85%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.95 (d, J=4.6, 3H, NHCH₃), 5.17 (s,2H, NH₂), 6.45 (d, J=5.4, 1H, H_(Py)), 6.80-6.88 (m, 2H, H_(arom)), 7.02(m, 1H, H_(arom)), 8.03 (br q, J=4.6, 1H, NHCH₃), 8.31 (s, 1H,H_(arom)), 8.48 (d, J=5.4, 1H, H_(Py));

¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 27.1, 104.0, 108.7 (d, J_(FC)=21.2),116.4 (d, J_(FC)=5.6), 117.0 (d, J_(FC)=21.2), 121.9, 134.3 (d,J_(FC)=12.9), 139.2 (br), 143.2 (d, J_(FC)=9.3), 150.2 (d, J_(FC)=240),152.7, 153.1, 155.3, 160.1; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −131.3; LC-MS(m/z): 286.1 (M+H, 100), rt=1.87 min.

Synthesis 612-fluoro-4-(3-(4-methylpiperazin-1-yl)pyrido[3,2-b]pyrazin-8-yloxy)aniline

Method E1 was used with Tert-butyl2-fluoro-4-(3-(4-methylpiperazin-1-yl)pyrido[3,2-b]pyrazin-8-yloxy)phenylcarbamate(125 mg, 0.28 mmol) to give the title compound as a yellow solid. Yield:76 mg (75%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.24 (s, 3H, CH₃), 2.46 (m, 4H,N(CH₂CH₂)₂NMe), 3.84 (m, 4H, N(CH₂CH₂)₂NMe), 5.16 (s, 2H, NH₂), 6.49 (d,J=5.3, 1H, H_(Py)), 6.80 (m, 1H, H_(arom)), 6.86 (m, 1H, H_(arom)), 7.02(m, 1H, H_(arom)), 8.53 (d, J=5.3, 1H, H_(Py)), 8.83 (s, 1H, H_(arom));¹³C-NMR (DMSO-d₆), δ (ppm), J (HZ): 43.9, 45.7, 54.2, 104.4, 108.7 (d,J_(FC)=21.1), 116.4 (d, J_(FC)=5.8), 117.0 (d, J_(FC)=2.8), 122.1, 134.4(d, J_(FC)=12.8), 136.0, 143.2 (d, J_(FC)=9.4), 150.2 (d, J_(FC)=240),152.0, 153.5, 153.8, 161.6; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −131.3; LC-MS(0.67 min): m/z calcd. for C₁₈H₂₀FN₆O [M+H⁺]: 355.1. Found: 355.1.

Synthesis 628-(4-amino-3-(methylthio)phenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one

Using Method E1 with tert-butyl4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenylcarbamate(131 mg, 0.3 mmol), the title compound (97 mg, 99%) was obtained as apale yellow powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.37 (s, 3H, CH₃); 2.42 (s, 3H, CH₃);5.17 (bs, 2H, NH₂), 6.43 (d, 1H, H_(Py) J=5.6 Hz), 6.79 (d, 1H,H_(arom), J=8.6 Hz), 6.86 (dd, 1H, H_(arom), J=8.6 Hz, J=2.6 Hz), 7.04(d, 1H, H_(arom), J=2.6 Hz), 8.21 (d, 1H, H_(Py), J=5.6 Hz), 12.69 (s,1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 15.6, 20.3, 105.0, 114.8,117.1, 119.7, 120.8, 121.6, 143.7, 144.8, 145.3, 150.4, 156.2, 158.4,161.0. LC-MS (m/z): 315 (M+H, 100), rt=3.68 min.

Synthesis 638-(4-amino-3-(methylthio)phenoxy)-3-methylpyrido[2,3-b]pyrazin-2(1H)-one

Using Method E1 with tert-butyl4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenylcarbamate(200 mg, 0.48 mmol), the title compound (34 mg, 23%) was obtained as apale yellow powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.36 (s, 3H, CH₃); 2.37 (s, 3H, CH₃);5.19 (s, 2H, NH₂), 5.96 (d, 1H, H_(Py), J=5.5 Hz), 6.75 (d, 1H,H_(arom), J=8.4 Hz), 6.88 (dd, 1H, H_(arom), J=8.5 Hz, J=2.6 Hz), 7.06(d, 1H, H_(arom), J=2.6 Hz), 8.28 (d, 1H, H_(Py), J=5.5 Hz), 12.33 (s,1H, NH). LC-MS (m/z): 315 (M+H, 100), rt=1.86 min.

Synthesis 648-(4-aminonaphthalen-1-yloxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one

Method E1 was used with tert-butyl4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamateto afford the title compound as a slightly yellow solid (309 mg, 0.971mmol, Quantitative).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.49 (s, 3H, Me), 5.86 (bs, 2H, NH₂),6.21 (d, 1H, H_(arom), J=5.7 Hz), 6.72 (d, 1H, H_(arom), J=8.2 Hz), 7.14(d, 1H, H_(arom), J=8.1 Hz), 7.40-7.46 (m, 2H, H_(arom)), 7.59 (d, 1H,H_(arom), J=7.8 Hz), 8.12 (d, 1H, H_(Py), J=5.7 Hz), 8.17 (d, 1H,H_(arom), J=8.2 Hz), 12.73 (s, 1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J(Hz): 20.47 (Me), 104.90, 106.34, 117.05, 118.71, 120.74, 123.08,123.33, 124.41, 126.39, 126.56, 138.40, 143.32, 145.42, 150.38, 156.29,158.53, 161.56. LC-MS (m/z): 319 (M+H, 100).

Synthesis 658-(4-aminonaphthalen-1-yloxy)-3-methylpyrido[2,3-b]pyrazin-2(1H)-one

Method E1 was used with tert-butyl4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamateto afford the title compound as a slightly yellow solid (354 mg, 1.11mmol, 66%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.50 (s, 3H, Me), 5.85 (bs, 2H, NH₂),6.46 (d, 1H, H_(PY), J=5.4 Hz), 6.72 (d, 1H, H_(arom), J=8.1 Hz), 7.17(d, 1H, H_(arom), J=8.1 Hz), 7.41-7.46 (m, 2H, H_(arom)), 7.66 (d, 1H,H_(arom), J=7.4 Hz), 8.15-8.17 (m, 2H, H_(arom)), 12.55 (s, 1H, NH).¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 20.93 (Me), 106.20, 107.66, 118.38,118.76, 121.00, 122.97, 123.33, 124.45, 126.32, 126.52, 138.28, 143.37,143.57, 144.89, 153.31, 154.48, 163.78. LC-MS (m/z): 319 (M+H, 100).

Synthesis 662-amino-8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-3(4H)-one

Method E1 was used with tert-butyl4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-to afford the title compound as a slightly pink solid (207 mg, 0.648mmol, 75%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.76 (bs, 2H, NH₂), 6.14 (d, 1H,H_(arom), J=5.6 Hz), 6.70 (d, 1H, H_(arom), J=8.1 Hz), 7.07 (d, 1H,H_(arom), J=8.1 Hz), 7.39-7.44 (m, 2H, H_(arom)), 7.62 (dd, 1H,H_(arom), J=7.5 Hz, J=2.0 Hz), 7.80 (d, 1H, H_(Py),6, J=5.6 Hz), 8.14(d, 1H, H_(arom), J=7.5 Hz), 12.50 (s, 1H, NH). ¹³C-NMR (DMSO-d₆), δ(ppm), J (Hz): 105.15, 106.46, 118.43, 118.59, 120.93, 122.97, 123.42,124.32, 126.15, 126.78, 139.07, 142.66, 142.84, 143.56, 151.49, 152.72,158.57. LC-MS (m/z): 320 (M+H, 100).

Synthesis 678-(4-amino-3-fluorophenoxy)-N,N-dimethylpyrido[3,2-b]pyrazin-3-amine

Method E1 was used with tert-butyl4-(3-(dimethylamino)pyrido[3,2-b]pyrazin-8-yloxy)-2-fluorophenylcarbamateto give the crude product (5% TBAF) as a beige solid, which was used insubsequent steps. Yield: 128 mg (78%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 3.27 (s, 6H, N(CH₃)₂), 5.15 (s, 2H,NH₂), 6.47 (d, J=5.2, 1H, H_(Py)), 6.80-6.88 (m, 2H, H_(arom)), 7.01 (m,1H, H_(arom)), 8.52 (d, J=5.2, 1H, H_(Py)), 8.69 (s, 1H, H_(arom))_(;)¹⁹F-NMR (DMSO-d₆), δ (ppm): −131.3; LC-MS (m/z): 300.1 (M+H, 100),rt=1.29 min.

Synthesis 683-amino-8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-2(1H)-one

Method E1 was used with tert-butyl4-(3-amino-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamateto afford the title compound as a slightly pink solid (198 mg, 0.620mmol, 60%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.80 (bs, 2H, NH₂), 6.14 (d, 1H,H_(arom), J=5.5 Hz), 6.70 (d, 1H, H_(arom), J=8.2 Hz), 7.13 (d, 1H,H_(arom), J=8.1 Hz), 7.42-7.44 (m, 2H, H_(arom)), 7.68 (d, 1H, H_(arom),J=8.2 Hz), 7.93 (d, 1H, H_(Py), J=5.5 Hz), 8.15 (d, 1H, H_(arom), J=7.2Hz), 12.28 (s, 1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 103.72,106.23, 113.28, 118.67, 121.16, 122.88, 123.32, 124.38, 126.16, 126.78,138.63, 143.06, 144.24, 146.42, 150.93, 152.39, 154.68. LC-MS (m/z): 320(M+H, 100).

Synthesis 698-(4-aminonaphthalen-1-yloxy)-2-(trifluoromethyl)pyrido[2,3-b]pyrazin-3(4H)-one

Method E1 was used with tert-butyl4-(3-oxo-2-(trifluoromethyl)-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamateto afford the title compound as a slightly yellow solid (56 mg, 0.150mmol, 66%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.91 (bs, 2H, NH₂), 6.30 (d, 1H,H_(Py), J=5.7 Hz), 6.73 (d, 1H, H_(arom), J=8.2 Hz), 7.19 (d, 1H,H_(arom), J=8.2 Hz), 7.41-7.48 (m, 2H, H_(arom)), 7.59 (d, 1H, H_(arom),J=8.5 Hz), 8.19 (d, 1H, H_(arom), J=8.0 Hz), 8.32 (d, 1H, H_(Py), J=5.7Hz), 13.46 (s, 1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 105.28,106.21, 116.39, 118.80, 120.63, 121.09, 123.14, 123.28, 124.50, 126.31,126.55, 137.91, 142.76 (CF₃), 143.69, 146.74, 153.37, 154.68, 163.25.LC-MS (m/z): 373 (M+H, 100).

Synthesis 69A8-(4-aminonaphthalen-1-yloxy)-3-(trifluoromethyl)pyrido[2,3-b]pyrazin-2(1H)-one

Method E1 was used with tert-butyl4-(2-oxo-3-(trifluoromethyl)-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamateto afford the title compound as a yellow solid (222 mg, 0.596 mmol,53%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.89 (bs, 2H, NH₂), 6.57 (d, 1H,H_(arom), J=5.2 Hz), 6.73 (d, 1H, H_(arom), J=8.1 Hz), 7.17 (d, 1H,H_(arom), J=8.1 Hz), 7.41-7.47 (m, 2H, H_(arom)), 7.67 (d, 1H, H_(arom),J=7.8 Hz), 8.17 (d, 1H, H_(arom), J=7.7 Hz), 6.26 (d, 1H, H_(arom),J=5.2 Hz), 13.54 (s, 1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz):118.80, 119.18, 121.09, 121.38, 123.12, 123.47, 124.58, 126.43, 126.49,131.55, 138.40, 141.98, 143.55, 146.04, 153.36, 155.01. LC-MS (m/z): 373(M+H, 100).

(VI) Ureas from Common Intermediates Synthesis 701-(4-chloro-3-(trifluoromethyl)phenyl)-3-(4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea (AA-042)

Method F1 (one pot deprotection of Boc and coupling with isocyanate):Tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate (0.240g, 0.67 mmol) is dissolved in trifluoroacetic acid (2 ml) and thesolution is stirred at room temperature under Argon atmosphere for 2 h.The solvent is evaporated under reduced pressure and the resulting darkoil is dissolved in THF (3 ml) and triethylamine (1 ml).1-chloro-4-isocyanato-2-(trifluoromethyl)benzene (0.180 g, 0.80 mmol) isadded in one portion, and the solution was stirred overnight at 45 Cunder Ar atmosphere. The solution is then cooled and evaporated and thecrude was crystallised from dichloromethane and diethyl ether to affordthe title compound (15 mg, 5% yield) as a brown solid.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.82 (d, 1H, J=5.5 Hz), 7.24 (d, 2H,J=8.9 Hz), 7.62 (d, 1H, J=9.0 Hz), 7.66 (dd, 1H, J=9.0, 2.6 Hz), 7.71(d, 2H, J=8.9 Hz), 8.12 (d, 1H, J=2.6 Hz), 8.34 (d, 1H, J=5.5 Hz), 8.41(s, 1H), 8.98 (bs, 1H), 9.18 (bs, 1H), 12.54 (bs, 1H); LC-MS (m/z): 476(M+H, 100).

Synthesis 711-(4-chloro-3-(trifluoromethyl)phenyl)-3-(4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-020)

Method E1 was used with tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate and4-chloro-3-trifluoromethylphenyl isocyanate to obtain the tithe compound(yield 83%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.53 (d, 1H, J=5.6 Hz), 7.18 (d, 2H,J=8.8 Hz), 7.58-7.70 (m, 4H), 8.13 (d, 1H, 1.9 Hz), 8.19 (s, 1H), 8.34(d, 1H, J=5.6 Hz), 9.14 (bs, 1H), 9.36 (bs, 1H), 12.88 (bs, 1H). LC-MS(m/z): 476 (M+H, 100).

Synthesis 721-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-016)

Method E1 was used with Tert-butyl4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate and3-tert-butyl-5-isocyanato-1-p-tolyl-1H-pyrazole to obtain the titlecompound (yield 55%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.25 (s, 3H), 1.28 (s, 9H), 6.35 (s,1H), 6.52 (d, 1H, J=5.4 Hz), 7.17 (d, 2H, J=9.0 Hz), 7.32-7.43 (ABsystem, 4H), 7.51 (d, 2H, J=9.0 Hz), 8.17 (s, 1H), 8.32 (d, 1H, 5.4 Hz),8.34 (bs, 1H), 9.12 (bs, 1H), 12.87 (bs, 1H); LC-MS (m/z): 510 (M+H,100).

Synthesis 731-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-040)

Method E1 was used with tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate and3-tert-butyl-5-isocyanato-1-p-tolyl-1H-pyrazole to obtain the titlecompound (yield 64%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.27 (s, 3H), 1.29 (s, 9H), 5.41 (s,1H), 6.07 (d, 1H, 5.8 Hz), 7.26 (d, 2H, 8.8 Hz), 7.32-7.41 (AB system,4H), 7.45 (d, 2H, J=8.8 Hz), 7.54 (d, 1H, 5.8 Hz), 8.28 (s, 1H), 8.30(bs, 1H), 9.0 (bs, 1H), 10.12 (bs, 1H). LC-MS (m/z): 510 (M+H, 100).

Synthesis 741-(4-chloro-3-(trifluoromethyl)phenyl)-3-(4-(2,3-dioxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-050)

Method F2 was used with8-(4-aminophenoxy)pyrido[2,3-b]pyrazine-2,3(1H,4H)-dione and4-chloro-3-trifluoromethylphenyl isocyanate to obtain the tithe compound(yield 38%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.62 (d, 1H, J=5.3 Hz), 7.41 (d, 2H,J=8.6 Hz), 7.52 (d, 2H, J=8.6 Hz), 8.26 (d, 1H, J=5.3 Hz), 9.03 (bs,1H), 9.41 (bs, 1H), 12.39 (bs, 1H), 12.98 (bs, 1H). LC-MS (m/z): 492(M+H, 100).

Synthesis 751-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-012)

Method F1 was used with tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamateand 2-fluoro-5-trifluoromethylphenyl isocyanate to obtain the titlecompound (yield 69%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.32 (d, 1H, J=4.8 Hz), 7.38-7.62 (m,6H), 7.88-7.94 (AB system, 2H), 8.06 (s, 1H), 8.18 (d, 1H, J=8.0 Hz),8.43 (d, 1H, J=4.8 Hz), 8.64 (bs, 1H), 10.52 (bs, 1H), 10.93 (bs, 1H).LC-MS (m/z): 510 (M+H, 100).

Synthesis 761-(4-chloro-3-(trifluoromethyl)phenyl)-3-(4-(1-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-037)

Method F1 was used with tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamateand 4-chloro-3-trifluoromethylphenyl isocyanate to obtain the titlecompound (yield 87%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.23 (d, 1H, J=5.9 Hz), 7.23 (d, 1H,J=8.0 Hz), 7.53-7.59 (m, 3H), 7.79 (d, 1H, 8.0 Hz), 7.84 (d, 2H, 8.5Hz), 8.08 (s, 1H), 8.21 (d, 1H, 6.8 Hz), 8.33 (d, 1H, 2.9 Hz), 8.38 (d,1H, 8.5 Hz), 11.46 (s, bs, 1H), 12.39 (bs, 1H). LC-MS (m/z): 526 (M+H,100).

Synthesis 771-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-033)

Method F1 was used with tert-butyl4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamateand 3-tert-butyl-5-isocyanato-1-p-tolyl-1H-pyrazole to obtain the titlecompound (yield 40%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.4 (m, 2H) 7.35-7.66 (m, 4H), 7.87(d, 1H, 8.5 Hz), 7.97 (d, 1H, 8.5 Hz), 8.12 (d, 1H, 8.7 Hz), 8.23 (s,1H), 8.27 (d, 1H, 5.7 Hz), 8.80 (bs, 1H), 9.18 (bs, 1H), 12.83 (bs, 1H).LC-MS (m/z): 560 (M+H, 100).

Synthesis 781-(4-chloro-3-(trifluoromethyl)phenyl)-3-(4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-013)

Method F1 was used with tert-butyl4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-ylcarbamateand 4-chloro-3-trifluoromethylphenyl isocyanate to obtain the titlecompound (yield 91%).

¹H-NMR (CD3OD), δ (ppm), J (Hz): 6.72 (d, 1H, J=5.7 Hz), 7. 46 (d, 1H,J=7.4 Hz), 7.42-7.63 (m, 4H), 7.82 (d, 1H, J=7.4 Hz), 7.91 (s, 1H), 7.99(d, 1H, J=3.0 Hz), 8.20 (d, 1H, J=8.3 Hz), 8.27 (d, 1H, J=5.7 Hz), 8.51(bs, 1H). LC-MS (m/z): 526 (M+H, 100).

Synthesis 791-(4-chloro-3-(trifluoromethyl)phenyl)-3-(2-(methylthio)-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-023)

Using Method F2 with8-(4-amino-3-(methylthio)phenoxy)pyrido[3,2-b]pyrazin-3(4H)-one and4-chloro-3-trifluoromethylphenyl isocyanate, the title compound (48 mg,92%) was obtained as a pale white powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.47 (s, 3H, CH₃); 6.61 (d, 1H,H_(Py) J=5.6 Hz), 7.07 (dd, 1H, H_(arom), J=8.8 Hz, J=2.6 Hz), 7.26 (d,1H, H_(arom), J=2.6 Hz), 7.62 (m, 2H, H_(arom)), 7.86 (d, 1H, H_(arom),J=8.7 Hz), 8.11 (m, 1H, H_(arom)), 8.18 (s, 1H, NH or CH), 8.21 (s, 1H,NH or CH), 8.36 (d, 1H, H_(Py) J=5.6 Hz), 9.75 (s, 1H, NH or CH), 12.89(s, 1H, NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 15.6, 106.1, 116.4,117.9, 118.2, 119.7, 121.6, 122.7, 123.4, 123.7, 124.2, 126.6, 131.9,133.6, 139.2, 145.3, 150.0, 150.9, 152.0, 152.4, 156.4, 160.7. LC-MS(m/z): 522 (M+H, 100), rt=5.24 min.

Synthesis 801-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(2-fluoro-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea (AA-023)

Method F2: A solution of8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-2(1H)-one (21.4 mg, 78.6μmol) in dry DMSO (1 mL) under Ar was treated with2-fluoro-5-trifluoro-phenylisocyanate (11.5 μL, 80 μmol) and the paleyellow solution was stirred at RT. After 3 h, the solution was dilutedwith H₂O (20 mL) and the precipitate was isolated by filtration.Stripping with toluene (3×20 mL) furnished the title compound as a beigepowder. Yield: 30 mg (81%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.94 ppm (d, 1H, J=5.5 Hz, H_(Py)),7.09 (m, 1H, H_(arom))), 7.32 (m, 1H, H_(arom)), 7.40 (m, 1H, H_(arom)),7.50 (m, 1H, H_(arom)), 8.23 (t, J=8.1 Hz, H_(arom)), 8.37 (d, J=5.5 Hz,H_(Py)), 8.40 (s, 1H, H_(arom)), 8.63 (m, 1H, H_(arom)), 9.23 (s, 1H,NH), 9.38 (s, 1H, NH), 12.58 (br s, 1H, NHAr); ¹³C-NMR (DMSO-d₆), δ(ppm), J (Hz): 108.5, 110.4, (d, J_(FC)=22,) Hz 116.1 (d, J_(FC)=21 Hz),116.5 (m), 119.5 (br), 121.9, 122.8, 124.6 (d, J_(FC)=11 Hz), 125.0,125.4 (d, J_(FC)=30 Hz), 128.5, 144.4 (br), 145.3, 148.7 (d, J_(FC)=10Hz), 151.4, 152.0, 152.3 (br), 152.4 (d, J_(FC)=246 Hz), 153.5 (d,J_(FC)=248 Hz), 155.0, 155.4 (br); ¹⁹F NMR (470 MHz, DMSO-d₆): δ=−60.7,−123.9, −125.2 ppm; LC-MS (m/z): 478.1 (M+H, 100), rt=4.89 min; HRMS(3.38 min): m/z calcd. for C₂₁H₁₃F₅N₅O₃ [M+H⁺]: 478.09331. Found:478.09355.

Synthesis 811-(4-chloro-3-(trifluoromethyl)phenyl)-3-(2-(methylthio)-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-045)

Using Method F2 with8-(4-amino-3-(methylthio)phenoxy)pyrido[2,3-b]pyrazin-2(1H)-one and4-chloro-3-trifluoromethylphenyl isocyanate, the title compound (15 mg,29%) was obtained as a pale brown powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.47 (s, 3H, CH₃); 6.90 (d, 1H,H_(Py) J=5.3 Hz), 7.10 (dd, 1H, H_(arom), J=8.8 Hz, J=2.4 Hz), 7.29 (d,1H, H_(arom), J=2.4 Hz), 7.63 (m, 2H, H_(arom)), 7.87 (d, 1H, H_(arom),J=8.8 Hz), 8.11 (m, 1H, H_(arom)), 8.22 (s, 1H, NH or CH), 8.36 (m, 1H,H_(Py)), 8.42 (s, 1H, NH or CH), 9.76 (s, 1H, NH or CH), 12.57 (s, 1H,NH). ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 15.7, 110.0, 116.4, 118.0,119.9, 122.2, 122.7, 123.4, 124.1, 126.7, 131.6, 131.9, 133.7, 138.8,139.2, 149.8, 152.4. LC-MS (m/z): 522 (M+H, 100), rt=5.10 min.

Synthesis 821-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(2-(methylthio)-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-024)

Using Method F2 with8-(4-amino-3-(methylthio)phenoxy)pyrido[3,2-b]pyrazin-3(4H)-one and2-fluoro-5-trifluoromethylphenyl isocyanate, the title compound (26 mg,62%) was obtained as a powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.48 (s, 3H, CH₃), 6.61 (d, 1H,H_(Py, 5), J=5.6 Hz), 7.06 (dd, 1H, H_(arom), J=8.6 Hz, J=2.3 Hz), 7.24(d, 1H, H_(arom), J=2.3 Hz), 7.39 (m, 1H, H_(arom)), 7.49 (m, 1H,H_(arom)), 7.85 (d, 1H, H_(arom), J=8.7 Hz), 8.18 (s, 1H, NH or CH),8.36 (d, 1H, H_(Py,6), J=5.6 Hz), 8.64 (m, 1H, H_(arom)), 8.68 (s, 1H,NH or CH), 9.53 (s, 1H, NH or CH), 12.90 (s, 1H, NH). ¹³C-NMR (DMSO-d₆),δ (ppm), J (Hz): 15.3, 106.1, 115.9, 116.1, 116.7, 117.6, 118.2, 119.2,122.7, 125.1, 125.4, 128.6, 132.2, 133.2, 145.4, 150.1, 150.9, 152.0,152.4, 154.4, 156.3, 160.7. LC-MS (m/z): 506 (M+H, 100), rt=4.85 min.

Synthesis 831-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(2-(methylthio)-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-046)

Using Method F2 with8-(4-amino-3-(methylthio)phenoxy)pyrido[2,3-b]pyrazin-2(1H)-one and2-fluoro-5-trifluoromethylphenyl isocyanate, the title compound (37 mg,73%) was obtained as a powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.48 (s, 3H, CH₃), 6.89 (d, 1H,H_(Py, 5), J=5.3 Hz), 7.09 (dd, 1H, H_(arom), J=8.8 Hz, J=2.5 Hz), 7.26(d, 1H, H_(arom), J=2.5 Hz), 7.39 (m, 1H, H_(arom)), 7.50 (m, 1H,H_(arom)), 7.85 (d, 1H, H_(arom), J=8.8 Hz), 8.36 (d, 1H, H_(Py,6),J=5.2 Hz), 8.42 (s, 1H, NH or CH), 8.64 (m, 1H, H_(arom)), 8.69 (s, 1H,NH or CH), 9.54 (s, 1H, NH or CH), 12.60 (s, 1H, NH). ¹³C-NMR (DMSO-d₆),δ (ppm), J (Hz): 15.4, 110.0, 115.9, 116.1, 116.7, 117.7, 119.1, 119.4,122.7, 124.8, 125.1, 125.4, 128.5, 132.0, 133.3, 145.3, 149.9, 152.4,154.5. LC-MS (m/z): 506 (M+H, 100), rt=5.00 min.

Synthesis 841-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(2-(methylthio)-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-017)

Using Method F2 with8-(4-amino-3-(methylthio)phenoxy)pyrido[3,2-b]pyrazin-3(4H)-one and-tert-butyl-5-isocyanato-1-tolyl-1H-pyrazole, the title compound (5 mg,8%) was obtained as a white powder after purification on silica gel(Eluent: DCM/EtOAc: 1/1, Rf=0.57).

¹H-NMR (CDCl₃), δ (ppm), J (Hz): 1.31 (s, 9H, tert-Bu), 2.23 (s, 3H,CH₃), 2.31 (s, 3H, SCH₃), 6.30 (s, 1H), 6.36 (s, 1H), 6.49 (d, 1H,H_(Py) J=5.8 Hz), 7.02 (dd, 1H, H_(arom), J=8.9 Hz, J=2.7 Hz), 7.19 (m,4H, H_(arom)), 7.31 (d, 1H, H_(arom), J=8.3 Hz), 7.81 (s, 1H, NH or CH),8.16 (d, 1H, H_(arom), J=8.9 Hz), 8.26 (s, 1H, NH or CH), 8.30 (d, 1H,H_(Py), J=5.8 Hz), 11.37 (s, 1H, NH). LC-MS (m/z): 556 (M+H, 100).

Synthesis 851-(4-Chloro-3-(trifluoromethyl)phenyl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea (AA-025)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one and3-trifluoromethyl-4-chloro-phenylisocyanate to afford the titlecompound, yield: 88%.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.67 ppm (d, 1H, J=5.5 Hz, H_(Py)),7.06 (d, 1H, H_(arom)), 7.30 (d, 1H, H_(arom)), 7.66 (m, 2H, H_(arom)),8.12 (m, 2H, H_(arom)), 8.17 (s, 1H, H_(arom)), 8.38 (d, J=5.5 Hz,H_(Py)), 8.88 (s, 1H, NH), 9.92 (s, 1H, NH), 12.91 (br s, 1H, NHAr); ¹⁹FNMR (470 MHz, DMSO-d₆): δ=−61.5, −124.2 ppm; LC-MS (m/z): 494.1 (M+H,100), rt=5.24 min; HRMS (6.17 min): m/z calcd. for C₂₁H₁₃ClF₄N₆O₃[M+H⁺]: 494.06376. Found: 494.06335.

Synthesis 861-(2-Fluoro-5-(trifluoromethyl)phenyl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea (AA-026)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one and2-fluoro-5-trifluoromethyl-phenylisocyanate to afford the titlecompound, yield=80%.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.67 ppm (d, 1H, J=5.5 Hz, H_(Py)),7.08 (m, 1H, H_(arom)), 7.34 (m, 1H, H_(arom)), 7.40 (m, 1H, H_(arom)),7.51 (m, 1H, H_(arom)), 8.17 (s, 1H, H_(arom)), 8.23 (t, J=8.1 Hz,H_(arom)) 8.38 (d, J=5.5 Hz, H_(Py)), 8.64 (m, 1H, H_(arom)), 9.20 (s,1H, NH), 9.35 (s, 1H, NH), 12.91 (br s, 1H, NHAr); ¹⁹F NMR (470 MHz,DMSO-d₆): δ=−60.8, −124.0, 125.2 ppm; LC-MS (m/z): 478.1 (M+H, 100),rt=5.04 min; HRMS (3.38 min): m/z calcd. for C₂₁H₁₃F₅N₅O₃ [M+H⁺]:478.09331. Found: 478.09355.

Synthesis 871-(4-Chloro-3-(trifluoromethyl)phenyl)-3-(2-fluoro-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea (AA-048)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-2(1H)-one and3-trifluoromethyl-4-chloro-phenylisocyanate to afford the title compoundas a beige powder. Yield: 60 mg (79%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.93 ppm (d, J=5.5 Hz, 1H, H_(Py)),7.08 (m, 1H, H_(arom)), 7.30 (m, 1H, H_(arom)), 7.64 (m, 2H, H_(arom)),8.12 (m, 2H, H_(arom)), 8.37 (d, J=5.5 Hz, 1H, H_(Py)), 8.41 (s, 1H,H_(arom)), 8.78 (s, 1H, NH), 9.57 (s, 1H, NH), 12.58 (br s, 1H, NHAr);¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 116.5, 108.6 110.4, ppm (d,J_(FC)=23 Hz), 154.9 (br), 116.6 (d, J_(FC)=6 Hz), 121.7, 122.6, 123.0,123.5, 123.8, 124.6 (d, J_(FC)=11 Hz), 125.3, 126.0, 126.8 (qu,J_(FC)=30 Hz), 128.5, 132.1, 139.1, 144.4 (br), 145.3, 148.9 (d,J_(FC)=10 Hz), 152.2, 152.8 (d, J_(FC)=248 Hz); ¹⁹F NMR (470 MHz,DMSO-d₆): δ=−61.5, −125.0 ppm; LC-MS (m/z): 494.1 (M+H, 100), rt=4.89min; HRMS (3.38 min): m/z calcd. for C₂₁H₁₃F₅N₅O₃ [M+H⁺]: 478.09331.Found: 478.09355.

Synthesis 881-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-018)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-p-tolyl-1H-pyrazole to afford the titlecompound as an off-white solid. Yield: 35 mg (42%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.28 ppm (s, 9H, tert-Bu 2.40 (s, 3H,CH₃), 6.39 (s, 1H, H_(Py)), 6.66 (d, J=5.6 Hz, 1H, _(Py)rH), 7.41-7.29(m, 5H, H_(arom)), 7.06 (m, 1H, H_(arom)), 8.21-8.17 (m, 2H, H_(arom)),8.38 (d, J=5.6 Hz, 1H, H_(Py)), 8.79 (s, 1H, NH), 9.00 (s, 1H, NH),12.93 (br s, 1H, NH_(arom)); ¹⁹F NMR (470 MHz, DMSO-d₆): δ=−125.2 ppm;LC-MS (m/z): 528.1 (M+H, 100), rt=5.07 min; HRMS (6.12 min): m/z calcd.for C₂₈H₂₆FN₇NaO₃ [M+Na⁺]: 514.09090. Found: 514.09051.

Synthesis 891-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-019)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound. Yield: 50 mg (60%) of a cream colored solid.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.28 ppm (s, 9H, tert-Bu), 6.40 (s,1H, H_(Pyr)), 6.66 (d, J=5.6 Hz, 1H, H_(Py)), 7.04 (m, 1H, H_(arom)),7.29 (m, 1H, H_(arom)), 7.42 (m, 1H, H_(arom)), 7.55-7.53 (m, 4H,H_(arom)), 8.17-8.16 (m, 2H, H_(arom)), 8.37 (d, J=5.6 Hz, 1H, H_(Py)),8.83 (s, 1H, NH), 8.98 (s, 1H, NH), 12.90 (br s, 1H, NHAr); ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 30.2, 32.0, 95.1, 106.5, 108.5 (d, J_(FC)=22Hz), 116.4 (d, J_(FC)=3 Hz), 118.4, 121.8, 124.4, 124.9 (d, J_(FC)=12Hz), 127.4, 129.3, 136.9, 138.4, 145.5, 148.6 (d, J_(FC)=10 Hz), 151.2,151.3, 152.2, 152.3 (d, J_(FC)=245 Hz), 153.3, 156.4, 160.5, 160.8,171.2; ¹⁹F NMR (470 MHz, DMSO-d₆): δ=−125.2 ppm; LC-MS (m/z): 514.2(M+H, 100), rt=4.93 min; HRMS (5.95 min): m/z calcd. for C₂₇H₂₃FN₇O₃[M+H⁺]: 514.19974. Found: 514.19964.

Synthesis 901-(2-fluoro-4-(3-methyl-2-oxo-1,2-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-049)

Method F2 was used with8-(4-amino-3-fluorophenoxy)-3-methylpyrido[3,2-b]pyrazin-2(1H)-one and2-fluoro-5-trifluoromethylphenyl isocyanate to afford the titlecompound; yield=85%.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.49 ppm (s, 3H, CH₃) 6.89 (d, 1H,J=5.6 Hz, _(Py)rH), 7.08 (m, 1H, H_(arom)), 7.32 (m, 1H, H_(arom)), 7.41(m, 1H, H_(arom)), 7.53 (m, 1H, H_(arom)), 8.23 (t, 1H, H_(arom)), 8.33(d, J=5.6 Hz, 1H, H_(Py)), 8.64 (m, 1H, H_(arom)), 9.19 (s, 1H, NH),9.35 (s, 1H, NH), 12.42 (br s, 1H, NH_(arom))_(;) ¹³C-NMR (DMSO-d₆), δ(ppm), J (Hz): 28.0, 79.4, 106.8, 109.0, 116.0, 118.5, 123.9, 125.8,145.6, 150.5, 151.2, 152.2, 153.1, 156.4, 160.3; ¹⁹F NMR (470 MHz,DMSO-d₆): δ=−60.7, −124.0, −125.3 ppm; LC-MS (m/z): 492.1 (M+H, 100),rt=4.98 min; HRMS (6.04 min): m/z calcd. for C₂₂H₁₄F₅N₅NaO₃ [M+Na⁺]:514.09090. Found: 514.09051.

Synthesis 911-(2-Fluoro-5-(trifluoromethyl)phenyl)-3-(4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-ylthio)phenyl)urea(AA-027)

Method F2 was used with8-(4-aminophenylthio)pyrido[2,3-b]pyrazin-3(4H)-one (36.7 mg, 136 mmol)and 2-fluoro-5-trifluoromethyl-phenylisocyanate (22.5 μL, 156 136 μmol)to afford the title compound. Yield: 53 mg (82%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.40 (d, J=5.3, 1H, H_(Py)), 7.43 (m,1H, H_(arom)), 7.52 (m, 1H, H_(arom)), 7.60 (d, J=8.3, 2H, H_(arom)),7.70 (d, J=8.3, 2H, H_(arom)), 8.20-8.22 (m, 2H, H_(Py)), 8.62 (m, 1H,H_(arom)), 9.03 (d, ⁴J_(FH)=2.6, 1H, NH), 9.53 (s, 1H, NH), 12.87 (br s,1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 114.6, 116.2 (d,J_(FC)=21.1), 116.8 (m), 119.6, 119.7, 122.8, 123.0, 125.1 (d,J_(FC)=40), 125.4 (m), 128.4 (d, J_(FC)=11.1), 136.9, 141.2, 143.4,150.0, 151.0, 152.0, 152.4, 153.6 (d, J_(FC)=21.1), 156.8; LC-MS (m/z):476.0 (M+H, 100), rt=5.42 min; HRMS (6.53 min): m/z calcd. forC₂₁H₁₄F₄N₅O₂S [M+H⁺]: 476.07988. Found: 476.07980.

Synthesis 921-(2-Fluoro-4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-028)

Method F2 was used with 2-fluoro-5-trifluoromethyl-phenylisocyanate and8-(4-amino-3-fluorophenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one toafford the title compound, yield 81%.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.43 (s, 3H, CH₃), 6.60 (d, J=5.4,1H, H_(Py)), 7.07 (m, 1H, H_(arom)), 7.33 (m, 1H, H_(arom)), 7.41 (m,1H, H_(arom)), 7.51 (m, 1H, H_(arom)), 8.24 (m, 1H, H_(arom)), 8.29 (m,1H, H_(arom)), 8.64 (d, J=5.4, 1H, H_(Py)), 9.20 (s, 1H, NH), 9.35 (s,1H, NH); ¹³C NMR (126 MHz, DMSO-d₆): δ=160.3, 156.4, 153.1, 152.2,151.2, 150.5, 145.6, 125.8, 123.9, 118.5, 116.0, 109.0, 106.8, 79.4,28.0 ppm; ¹⁹F NMR (470 MHz, DMSO-d₆): δ=−60.7, −124.0, −125.3 ppm; LC-MS(m/z): 492.1 (M+H, 100), 5.17 min; HRMS (7.15 min): m/z calcd. forC₂₂H₁₄F₅N₆O₃ [M+H⁺]: 492.10896. Found: 492.10843.

Synthesis 931-(3-Tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(4-(2,3-dioxo-1,2,3,4-tetrahydropyrido[3,2-b]pyrazin-8-yloxy)-2-fluorophenyl)urea(AA-091)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[3,2-b]pyrazine-2,3(1H,4H)-dione (50mg, 173 μmol) and a solution of3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole (5.7 mL of a 61 mMsolution in CH₂Cl₂, 347 μmol) to give the title compound as a whitesolid. Yield: 65 mg (71%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 6.38 (s, 1H,_(Py)razoleH), 6.57 (d, 1H, J=5.3, H_(Py)), 7.00 (m, 1H, H_(arom)), 7.22(m, 1H, H_(arom)), 7.42 (m, 1H, J=8.3, H_(arom)), 7.54 (m, 4H,H_(arom)), 7.96 (d, 1H, J=5.3, H_(Py)), 8.11 (m, 1H, H_(arom)), 9.05 (s,1H, NH), 9.10 (s, 1H, H_(arom)), 11.91 (br s, 1H, NH), 12.40 (br s, 1H,NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 30.2, 32.0, 95.7, 108.2 (d,J_(FC)=22.4), 112.5, 116.1, 121.8, 124.3, 124.6 (d, J_(FC)=10.7), 127.3,129.2, 136.9, 138.5, 140.6, 143.2, 148.7 (d, J_(FC)=9.8), 150.3, 151.6,152.3 (d, J_(FC)=245), 154.8, 156.0, 160.8; ¹⁹F-NMR (DMSO-d₆), δ (ppm):−124.4; LC-MS (m/z): 531.1 (M+H, 100), rt=2.54 min; HRMS (3.07 min): m/zcalcd. for C₂₇H₂₆FN₇O₄ [M+H⁺]: 530.19466. Found: 530.19433.

Synthesis 941-(3-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-086)

Using Method F2 with8-(4-amino-2-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one (50 mg, 0.18mmol), the title compound (42 mg, 49%) was obtained as a brown powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.60 (d, 1H, H_(Py), J=5.7 Hz),7.27-7.30 (m, 1H, H_(arom)), 7.44-7.49 (m, 2H, H_(arom)), 7.53-7.57 (m,1H, H_(arom)), 7.81 (dd, 1H, H_(arom), J=12.9 Hz, J=2.3 Hz), 8.24 (s,1H, NH or CH), 8.39 (d, 1H, H_(Py) J=5.7 Hz), 8.63 (dd, 1H, H_(arom),J=7.4 Hz and J=2.0 Hz), 9.04 (s, 1H, NH or CH), 9.54 (s, 1H, NH or CH),13.00 (s, 1H, NH). ¹⁹F-NMR (δ, ppm, DMSO-d₆): −60.06, −123.20, −128.06.LC-MS (m/z): 478 (M+H, 100), rt=2.65 min. HRMS (EI): m/z (M+H, 100)calcd for C₂₁H₁₂F₅N₅O₃. 478.0933. Found: 478.0929.

Synthesis 951-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(3-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-087)

Using Method F2 with8-(4-amino-2-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one (50 mg, 0.18mmol), the title compound (26 mg, 28%) was obtained as a brown powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.32 (s, 9H, tert-Bu), 6.43 (s, 1H,CH), 6.58 (d, 1H, H_(Py) J=5.5 Hz), 7.21-7.25 (m, 1H, H_(arom)), 7.37(t, 1H, H_(arom), J=9.0 Hz), 7.43-7.48 (m, 1H, H_(arom)), 7.56-7.59 (m,4H, H_(arom)), 7.74 (dd, 1H, H_(arom), J=13.2 Hz, J=2.0 Hz), 8.23 (s,1H, NH or CH), 8.38 (d, 1H, H_(Py), J=5.3 Hz), 8.57 (s, 1H, NH or CH),9.41 (s, 1H, NH or CH), 12.99 (s, 1H, NH). ¹⁹F-NMR (δ, ppm, DMSO-d6):−128.21. LC-MS (m/z): 514 (M+H, 100), rt=2.61 min. HRMS (EI): m/z (M+H,100) calcd for C₂₇H₂₄FN7O3: 514.1997. Found: 514.2001.

Synthesis 961-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(3-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-088)

Using Method F2 with8-(4-amino-2-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one (50 mg, 0.18mmol), the title compound (26 mg, 27%) was obtained as a brown powder.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.32 (s, 9H, tert-Bu), 2.41 (s, 3H,CH₃), 6.41 (s, 1H, CH), 6.58 (d, 1H, H_(Py), J=5.7 Hz), 7.21-7.24 (m,1H, H_(arom)), 7.35-7.40 (m, 3H, H_(arom)), 7.42-7.45 (m, 2H, H_(arom)),7.74 (dd, 1H, H_(arom), J=13.2 Hz, J=2.2 Hz), 8.23 (s, 1H, NH or CH),8.38 (d, 1H, H_(Py), J=5.7 Hz), 8.51 (s, 1H, NH or CH), 9.41 (s, 1H, NHor CH), 12.99 (s, 1H, NH). ¹⁹F-NMR (δ, ppm, DMSO-d6): −128.21. LC-MS(m/z): 528 (M+H, 100), rt=2.67 min. HRMS (EI): m/z (M+H, 100) calcd forC₂₈H₂₆FN703: 528.2153. Found: 528.2156.

Synthesis 971-(2-Fluoro-4-(3-morpholinopyrido[2,3-b]pyrazin-8-yloxy)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-054)

Method F3: 2-Fluoro-4-(3-morpholinopyrido[2,3-b]pyrazin-8-yloxy)aniline(29 mg, 85 mmol) was dissolved in dry THF (5 mL) to give a light yellowsolution. 2-Fluoro-5-trifluoromethyl-phenylisocyanate (25 μL, 170 μmol)was added to this solution and after 3 h, all volatiles were evaporated.The resulting yellow oil was dissolved in CH₂Cl₂ and purified by columnchromatography on silica. Elution with EtOAc gave the product as ayellow band. Yield: 44 mg (96%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 3.76 (m, 4H, N(CH₂CH₂)₂O), 3.84 (m,4H, N(CH₂CH₂)₂O), 6.70 (d, 1H, J=5.3, H_(Py)), 7.04 (m, 1H, H_(arom)),7.30 (m, 1H, H_(arom)), 7.41 (m, 1H, H_(arom)), 7.51 (m, 1H, H_(arom)),8.21 (m, 1H, H_(arom)), 8.62-8.65 (m, 2H, H_(arom)+H_(Py)), 8.84 (s, 1H,H_(arom)), 9.18 (s, 1H, NH), 9.35 (s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ(ppm), J (Hz): 44.4, 65.9, 105.9, 108.3 (d, J_(FC)=22.3), 116.1 (d,J_(FC)=20.7), 116.3, 116.6, 119.5, 122.6, 122.8, 124.2 (d, J_(FC)=10.7),125.0, 125.4 (m), 128.5 (d, J_(FC)=11.4), 136.3, 149.6 (d, J_(FC)=10.4),152.1 (d, J_(FC)=16.4), 152.4 (d, J_(FC)=245), 153.4 (d, J_(FC)=248),153.7, 153.9, 160.3; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −60.7, −124.0, −125.3;LC-MS (m/z): 547.0 (M+H, 100), rt=4.35 min; HRMS (6.65 min): m/z calcd.for C₂₅H₁₉F₅N₆O₃ [M+H⁺]: 547.15116. Found: 547.15163.

Synthesis 981-(2-fluoro-4-(3-(methylamino)pyrido[3,2-b]pyrazin-8-yloxy)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-055)

Method F2 was used with 2-fluoro-5-(trifluoromethyl)phenyl isocyanateand 8-(4-amino-3-fluorophenoxy)-N-methylpyrido[3,2-b]pyrazin-3-amine togive the title compound as a white solid. Yield: 56 mg (80%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.95 (d, J=4.6, 3H, NHCH₃), 6.61 (d,J=5.4, 1H, H_(Py)), 7.03 (m, 2H, H_(arom)), 7.28 (m, 1H, H_(arom)), 7.40(m, 1H, H_(arom)), 7.50 (m, 1H, H_(arom)), 8.03 (br q, J=4.6, 1H,NH_(Me)), 8.20 (m, 1H, H_(arom)), 8.31 (s, 1H, H_(arom)), 8.54 (d,J=5.4, 1H, H_(Py)), 8.64 (m, 1H, H_(arom)), 9.17 (s, 1H, NH), 9.34 (s,1H, NH);

¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 27.1, 105.3, 108.3 (d, J_(FC)=22.3),116.1 (d, J_(FC)=20.5), 116.3 (d, J_(FC)=2.6), 116.6 (m), 119.4 (m),122.0 (d, J_(FC)=2.3), 122.8, 125.0, 125.4 (m), 128.5 (d, J_(FC)=11.4),139.6 (br), 149.6 (d, J_(FC)=10.3), 152.0, 152.4 (d, J_(FC)=245), 152.8,153.4, 153.4 (d, J_(FC)=248), 155.3, 160.4; ¹⁹F-NMR (DMSO-d₆), δ (ppm):−60.8, −124.0, −125.3; LC-MS (m/z): LC-MS: m/z 491.0 (M+H, 100), rt=1.87min; HRMS (6.65 min): m/z calcd. for C₂₅H₁₉F₅N₆O₃ [M+H⁺]: 547.15116.Found: 547.15163.

Synthesis 991-(3-Tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-041)

Method F3 was used with 3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazoleand 8-(4-Amino-3-fluorophenoxy)-3-methylpyrido[2,3-b]pyrazin-2(1H)-one.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 2.49 (s, 3H,CH₃), 6.85 (s, 1H, H_(Py)), 6.85 (d, 1H, J=5.6 Hz, H_(Py)), 7.04 (m, 1H,H_(arom)), 7.27 (m, 1H, H_(arom)), 7.43 (m, 1H, H_(arom)), 7.54 (m, 4H,H_(arom)), 8.15 (m, 1H, H_(arom)), 8.31 (d, J=5.6 Hz, 1H, H_(Py)), 8.83(s, 1H, NH), 8.99 (s, 1H, NH), 12.40 (br s, 1H, NHAr); ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 21.0, 108.5 (d, J_(FC)=21), 109.7, 116.1 (d,J_(FC)=6), 116.6 (m), 119.5 (br), 121.8 (m), 122.8, 124.5 (d,J_(FC)=10.8), 125.0, 125.4 (m), 128.1, 128.5 (d, J_(FC)=11.4), 143.9,145.0, 148.8 (d, J_(FC)=10.4), 151.3, 152.0, 152.3 (d, J_(FC)=246),153.4 (d, J_(FC)=249), 154.5; ¹⁹F NMR (470 MHz, DMSO-d₆): δ=−125.3 ppmLC-MS (m/z): 528.1 (M+H, 100), rt=4.97 min; HRMS (6.04 min): m/z calcd.for C₂₂H₁₄F₅N₅NaO₃ [M+Na⁺]: 514.09090. Found: 514.09051.

Synthesis 1001-(2-Fluoro-4-(3-(4-methylpiperazin-1-yl)pyrido[3,2-b]pyrazin-8-yloxy)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-056)

Method F3 was used with 2-Fluoro-5-(trifluoromethyl)phenyl isocyanateand2-fluoro-4-(3-(4-methylpiperazin-1-yl)pyrido[3,2-b]pyrazin-8-yloxy)anilineto give the title compound as a yellow solid. Yield: 30 mg (64%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 2.26 (s, 3H, CH₃), 2.48 (m, 4H,N(CH₂CH₂)₂NMe), 3.86 (m, 4H, N(CH₂CH₂)₂NMe), 6.68 (d, J=5.4, 1H,H_(Py)), 7.06 (m, 2H, H_(arom)), 7.32 (m, 1H, H_(arom)), 7.43 (m, 1H,H_(arom)), 7.52 (m, 1H, H_(arom)), 8.22 (m, 1H, H_(arom)), 8.63 (d,J=5.4, 1H, H_(Py)), 8.66 (m, 1H, H_(arom)), 8.86 (s, 1H, H_(arom)), 9.19(s, 1H, NH), 9.36 (s, 1H, NH);

¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 43.9, 45.7, 54.2, 105.8, 108.3 (d,J_(FC)=22.3), 116.1 (d, J_(FC)=21.4), 116.3 (d, J_(FC)=2.6), 116.6 (m),119.5 (m), 122.0 (d, J_(FC)=2.3), 122.3, 125.0, 125.4 (m), 128.5 (d,J_(FC)=11.4), 136.4, 149.6, 149.7, 151.5, 152.4 (d, J_(FC)=245), 153.4,153.4 (d, J_(FC)=248), 153.6, 153.8, 160.3; ¹⁹F-NMR (DMSO-d₆), δ (ppm):−60.8, −124.0, −125.3 ppm; LC-MS (m/z): 560.1 (M+H, 100), rt=3.18 min;HRMS (6.65 min): m/z calcd. for C₂₅H₁₉F₅N₆O₃ [M+H⁺]: 547.15116. Found:547.15163.

Synthesis 10131-(3-Tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-(4-methylpiperazin-1-yl)pyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-053)

Method F3 was used with 3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazoleand2-fluoro-4-(3-(4-methylpiperazin-1-yl)pyrido[3,2-b]pyrazin-8-yloxy)anilineto give the title compound as a cream-colored solid. Yield: 44 mg (77%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 2.31 (br, 3H,CH₃), 2.56 (br, 4H, N(CH₂CH₂)₂NMe), 3.88 (m, 4H, N(CH₂CH₂)₂NMe), 6.41(s, 1H, H_(Pyz)), 6.66 (d, J=5.3, 1H, H_(Py)), 7.03 (m, 1H, H_(arom)),7.27 (m, 1H, H_(arom)), 7.45 (m, 1H, H_(arom)), 7.56 (m, 4H, H_(arom)),8.15 (m, 1H, H_(arom)), 8.61 (d, J=5.3, 1H, H_(Py)), 8.86 (m, 2H,NH+H_(arom)), 9.00 (s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz):30.2, 32.0, 43.7, 45.3, 54.0, 95.2, 105.8, 108.3 (d, J_(FC)=22.3), 116.2(d, J_(FC)=2.6), 121.8 (d, J_(FC)=2.3), 122.4, 124.4, 124.5 (d,J_(FC)=11.8), 127.3, 129.3, 136.4, 137.0, 138.5, 149.3 (d, J_(FC)=10.6),151.4, 152.2, 152.4 (d, J_(FC)=245), 153.6, 153.8, 160.3, 160.8; ¹⁹F-NMR(DMSO-d₆), δ (ppm): −60.8, −124.0, −125.3 ppm; LC-MS (m/z): 596.1 (M+H,100), rt=3.10 min; HRMS (6.65 min): m/z calcd. for C₂₅H₁₉F₅N₆O₃ [M+H⁺]:547.15116. Found: 547.15163.

Synthesis 1021-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-006)

Method F2 was used with 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-3(4H)-oneand 3-tert-butyl-5-isocyanato-1-p-tolyl-1H-pyrazole to afford the titlecompound as a white solid (46 mg, 65%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.26 (s, 9H, tert-Bu), 2.36 (s, 3H,Me), 6.32 (s, 1H, H_(arom)), 6.58 (d, 1H, H_(Py), J=6.6 Hz), 6.82 (d,1H, H_(arom), J=6.8 Hz), 7.21 (d, 1H, H_(arom), J=7.2 Hz), 7.30-7.43 (m,6H, H_(arom)), 8.14 (s, 1H, H_(arom),), 8.35 (d, 1H, H_(Py), J=6.8 Hz),8.74 (s, 1H, NH_(urea)), 9.30 (s, 1H, NH_(urea)), 12.88 (s, 1H,NH_(lactame)). ¹³C-NMR (δ, ppm, DMSO-d₆): 20.55 (CH₃), 30.17 (tert-Bu),31.96 (tert-Bu), 95.84, 99.49, 106.47, 109.65, 113.47, 115.03, 118.50,124.15 (2*C), 129.30, 129.57 (2*C), 130.48, 136.12, 136.59, 136.84,141.56, 151.00, 151.72, 152.06, 154.39, 160.41, 160.50. HRMS (EI): m/z[M+H] calcd for C₂₈H₂₇N₇O₃: 510.2248. Found: 510.2253.

Synthesis 1031-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-034)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-2(1H)-one and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound as a white solid (11 mg, 17%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.31 (s, 9H, tert-Bu), 6.44 (s, 1H,H_(arom)), 6.65 (d, 1H, H_(arom), J=5.4 Hz), 7.41-7.47 (m, 2H,H_(arom)), 7.57-7.62 (m, 5H, H_(arom)), 7.66-7.69 (m, 1H, H_(arom)),7.93 (d, 1H, H_(arom), J=8.4 Hz), 7.96 (d, 1H, H_(arom), J=8.3 Hz), 8.11(d, 1H, H_(arom), J=8.5 Hz), 8.27 (d, 1H, H_(arom), J=4.5 Hz), 8.47 (s,1H, H_(arom)), 8.82 (s, 1H, NH_(urea)), 9.15 (s, 1H, NH_(urea)), 12.82(s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 30.17(tert-Bu), 32.02 (tert-Bu), 95.74, 109.39, 111.11, 118.36, 121.83,122.27, 124.24 (2*C), 126.33, 126.76, 127.24, 129.28 (2*C), 132.25,137.19, 138.65, 144.91, 145.29, 152.31, 154.65, 160.81. HRMS (EI): m/z[M+H] calcd for C₃₁H₂₇N₇O₃: 546.2248. Found: 546.2248.

Synthesis 1041-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-038)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-2(1H)-one and1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound as a white solid (65 mg, 98%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.64 (d, 1H, H_(arom), J=5.3 Hz),7.38-7.42 (m, 2H, H_(arom)), 7.53 (t, 1H, H_(arom), J=8.9 Hz), 7.59 (t,1H, H_(arom), J=7.6 Hz), 7.70 (t, 1H, H_(arom), J=7.7 Hz), 7.94 (d, 1H,H_(arom), J=8.5 Hz), 8.07 (d, 1H, H_(arom), J=8.4 Hz), 8.26 (d, 1H,H_(arom), J=5.3 Hz), 8.28 (d, 1H, H_(arom), J=8.6 Hz), 8.44 (s, 1H,H_(arom)), 8.68 (d, 1H, H_(arom), J=7.2 Hz), 9.47 (s, 1H, NH_(urea)),9.51 (5, 1H, NH_(urea)), 12.77 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6),δ (ppm), J (Hz): 109.22, 115.97, 116.14, 116.66, 116.89, 118.09, 119.27,121.82, 122.07, 122.75, 123.01, 124.92, 125.18, 125.46, 126.29, 126.76,126.81, 127.54, 128.70, 128.79, 131.73. HRMS (EI): m/z [M+H] calcd forC₂₅H₁₅F₄N₅O₃: 510.1184. Found: 510.1180.

Synthesis 1051-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-014)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one and1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound as a slightly pink solid (50 mg, 61%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 2.54 (s, 3H, Me), 6.33 (d, 1H,H_(arom), J=5.7 Hz), 7.40-7.43 (m, 2H, H_(arom)), 7.54 (t, 1H, H_(arom),J=8.8 Hz), 7.60 (t, 1H, H_(arom), J=7.5 Hz), 7.72 (t, 1H, H_(arom),J=8.2 Hz), 7.88 (d, 1H, H_(arom), J=8.4 Hz), 8.10 (d, 1H, H_(arom),J=8.3 Hz), 8.19 (d, 1H, H_(arom), J=5.3 Hz), 8.27 (d, 1H, H_(arom),J=8.7 Hz), 8.70 (dd, 1H, H_(arom), J=7.3 Hz, J=2.0 Hz), 9.34 (s, 1H,NH_(urea)), 9.40 (s, 1H, NH_(urea)), 12.79 (s, 1H, NH_(lactame)).¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 20.51 (Me), 105.50, 115.97, 116.50,117.29, 117.98, 119.22, 121.50, 122.07, 122.72, 124.89, 125.29, 126.35,126.87, 127.38, 128.69, 131.71, 145.11, 145.67, 150.51, 152.39, 152.55,154.36, 156.33, 159.10, 160.59. HRMS (EI): m/z [M+H] calcd forC₂₆H₁₇F₄N₅O₃: 524.1340. Found: 524.1324.

Synthesis 1061-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-039)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)-3-methylpyrido[2,3-b]pyrazin-2(1H)-one and1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound as a slightly yellow solid (28 mg, 42%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 2.52 (s, 3H, Me), 6.60 (d, 1H,H_(arom), J=5.3 Hz), 7.39-7.41 (m, 2H, H_(arom)), 7.51-7.54 (m, 1H,H_(arom)), 7.60 (t, 1H, H_(arom), J=7.7 Hz), 7.71 (t, 1H, H_(arom),J=7.6 Hz), 7.95 (d, 1H, H_(arom), J=8.4 Hz), 8.07 (d, 1H, H_(arom),J=8.3 Hz), 8.22 (d, 1H, H_(arom), J=5.3 Hz), 8.26 (d, 1H, H_(arom),J=8.5 Hz), 8.68 (d, 1H, H_(arom), J=6.6 Hz), 9.39 (s, 1H, NH_(urea)),9.44 (s, 1H, NH_(urea)), 12.66 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6),δ (ppm), J (Hz): 20.97 (Me), 108.47, 115.95, 116.11, 116.58, 116.86,117.98, 119.20, 121.86, 121.97, 122.73, 124.90, 125.32, 126.30, 126.72,126.80, 127.06, 127.47, 128.68, 131.67, 143.93, 144.82, 145.02, 152.44,152.61, 154.41. HRMS (EI): m/z [M+H] calcd for C₂₆H₁₇F₄N₅O₃: 524.1340.Found: 524.1341.

Synthesis 1071-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-008)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound as a slightly yellow solid (65 mg, 80%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.29 (s, 9H, tert-Bu), 6.39 (d, 1H,H_(arom), J=5.7 Hz), 6.43 (s, 1H, H_(arom)), 7.38 (d, 1H, H_(arom),J=8.3 Hz), 7.44 (t, 1H, H_(arom), J=7.0 Hz), 7.55-7.61 (m, 5H,H_(arom)), 7.66 (t, 1H, H_(arom), J=7.6 Hz), 7.85 (d, 1H, H_(arom),J=8.4 Hz), 7.94 (d, 1H, H_(arom), J=8.3 Hz), 8.10 (d, 1H, H_(arom),J=8.6 Hz), 8.25 (s, 1H, H_(arom)), 8.27 (d, 1H, H_(arom), J=5.7 Hz),8.80 (s, 1H, NH_(urea)), 9.13 (s, 1H, NH_(urea)), 12.94 (s, 1H,NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 30.10 (tert-Bu),31.95 (tert-Bu), 95.68, 105.78, 117.03, 118.05, 118.40, 121.32, 122.35,124.16 (2*C), 126.22, 126.68, 126.92, 127.17, 127.71, 129.21 (2*C),132.12, 137.12, 138.58, 145.13, 145.44, 151.12, 152.10, 152.24, 156.46,160.74, 161.31. HRMS (EI): m/z [M+H] calcd for C₃₁H₂₇N₇O₃: 546.2248.Found: 546.2250.

Synthesis 1081-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-009)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound as a slightly pink solid (56 mg, 71%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.30 (s, 9H, tert-Bu), 2.48 (s, 3H,Me), 6.31 (d, 1H, H_(arom), J=5.6 Hz), 6.43 (s, 1H, H_(arom)), 7.37 (d,1H, H_(arom), J=8.3 Hz), 7.43-7.46 (m, 1H, H_(arom)), 7.55-7.67 (m, 6H,H_(arom)), 7.84 (d, 1H, H_(arom), J=8.3 Hz), 7.95 (d, 1H, H_(arom),J=8.3 Hz), 8.10 (d, 1H, H_(arom), J=8.6 Hz), 8.18 (d, 1H, H_(arom),J=5.6 Hz), 8.80 (s, 1H, NHurea), 9.12 (s, 1H, NHurea), 12.78 (s, 1H,NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 20.58 (CH₃), 30.13(tert-Bu), 31.98 (tert-Bu), 95.63, 105.52, 117.30, 117.33, 118.37,121.44, 122.36, 124.21 (2*C), 126.34, 126.68, 126.92, 127.21, 127.69,129.26 (2*C), 132.14, 137.15, 138.58, 145.08, 145.64, 150.55, 152.23,156.34, 159.15, 160.64, 160.75. HRMS (EI): m/z [M+H] calcd forC₃₂H₂₉N₇O₃: 560.2405. Found: 560.2407.

Synthesis 1091-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-035)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)-3-methylpyrido[2,3-b]pyrazin-2(1H)-one and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound as a white solid (50 mg, 41%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.29 (s, 9H, tert-Bu), 2.51 (3H, s,Me), 6.43 (d, 1H, H_(arom), J=5.3 Hz), 6.59 (s, 1H, H_(arom),), 7.38 (d,1H, H_(arom), J=8.3 Hz), 7.44 (t, 1H, H_(arom), J=7.3 Hz), 7.55-7.61 (m,5H, H_(arom)), 7.66 (t, 1H, H_(arom), J=7.6 Hz), 7.91-7.95 (m, 2H,H_(arom)), 8.09 (d, 1H, H_(arom), J=8.6 Hz), 8.21 (d, 1H, H_(arom),J=5.4 Hz), 8.80 (s, 1H, NH_(urea)), 9.13 (s, 1H, NH_(urea)), 12.65 (s,1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 20.95 (CH₃),30.09 (tert-Bu), 31.95 (tert-Bu), 95.66, 108.51, 116.89, 118.33, 118.81,121.79, 122.17, 124.16 (2*C), 126.28, 126.66, 126.69, 127.17, 127.74,129.21 (2*C), 132.04, 137.12, 138.58, 143.77, 144.88, 144.99, 152.15,152.24, 154.54, 160.74, 164.12. HRMS (EI): m/z [M+H] calcd forC₃₂H₂₉N₇O₃: 560.2405. Found: 560.2402.

Synthesis 1101-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-010)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-tolyl-1H-pyrazole to afford the titlecompound as a slightly yellow solid (80 mg, 70%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.29 (s, 9H, tert-Bu), 2.40 (s, 3H,Me), 6.40 (d, 1H, H_(arom), J=5.6 Hz), 6.41 (s, 1H, H_(arom)), 7.37-7.39(m, 3H, H_(arom)), 7.47 (d, 2H, H_(arom), J=8.1 Hz), 7.57 (t, 1H,H_(arom), J=7.6 Hz), 7.66 (t, 1H, H_(arom), J=7.6 Hz), 7.86 (d, 1H,H_(arom), J=8.4 Hz), 7.97 (d, 1H, H_(arom), J=8.3 Hz), 8.11 (d, 1H,H_(arom), J=8.6 Hz), 8.25 (s, 1H, H_(arom)), 8.26 (d, 1H, H_(arom),J=5.7 Hz), 8.77 (s, 1H, NH_(urea)), 9.13 (s, 1H, NH_(urea)), 12.94 (s,1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 20.51 (CH₃),30.11 (tert-Bu), 31.90 (tert-Bu), 95.01, 105.75, 117.02, 118.03, 118.17,121.30, 122.30, 124.26 (2*C), 126.21, 126.62, 126.89, 127.61, 129.61(2*C), 132.14, 136.05, 136.71, 137.09, 145.02, 145.44, 151.09, 152.08,156.44, 160.46, 161.30. HRMS (EI): m/z [M+H] calcd for C₃₂H₂₉N₇O₃:560.2405. Found: 560.2403.

Synthesis 1111-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-011)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-tolyl-1H-pyrazole to afford the titlecompound as a slightly pink solid (67 mg, 69%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.29 (s, 9H, tert-Bu), 2.40 (s, 3H,Me), 2.48 (s, 3H, Me), 6.31 (d, 1H, H_(arom), J=5.6 Hz), 6.41 (s, 1H,H_(arom)), 7.37-7.38 (m, 3H, H_(arom)), 7.47 (d, 2H, H_(arom), J=8.3Hz), 7.56 (t, 1H, H_(arom), J=7.5 Hz), 7.66 (t, 1H, H_(arom), J=7.4 Hz),7.84 (d, 1H, H_(arom), J=8.4 Hz), 7.97 (d, 1H, H_(arom), J=8.3 Hz), 8.11(d, 1H, H_(arom), J=8.6 Hz), 8.18 (d, 1H, H_(arom), J=5.6 Hz), 8.77 (s,1H, NH_(urea)), 9.13 (s, 1H, NH_(urea)), 12.80 (s, 1H, NH_(lactame)).¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 20.51 (2*CH₃), 30.11 (tert-Bu),31.91 (tert-Bu), 95.03, 105.51, 117.21, 117.33, 118.21, 121.40, 122.30,124.26 (2*C), 126.31, 126.60, 126.85, 127.60, 129.61 (2*C), 132.11,136.05, 136.71, 137.09, 145.04, 145.61, 150.48, 152.09, 156.28, 159.09,160.47, 160.60. HRMS (EI): m/z [M+H] calcd for C₃₃H₃₁N₇O₃: 574.2561.Found: 574.2558.

Synthesis 1121-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(3-methyl-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-036)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)-3-methylpyrido[2,3-b]pyrazin-2(1H)-one and3-tert-butyl-5-isocyanato-1-tolyl-1H-pyrazole to afford the titlecompound as a white solid (71 mg, 49%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.29 (s, 9H, tert-Bu), 2.40 (3H, s,Me), 2.52 (3H, s, Me), 6.41 (s, 1H, H_(arom),), 6.59 (d, 1H, H_(arom),J=5.4 Hz), 7.37-7.39 (m, 3H, H_(arom)), 7.47 (d, 2H, H_(arom), J=8.2Hz), 7.57 (t, 1H, H_(arom), J=7.6 Hz), 7.66 (t, 1H, H_(arom), J=7.6 Hz),7.92 (d, 1H, H_(arom), J=8.4 Hz), 7.96 (d, 1H, H_(arom), J=8.3 Hz), 8.09(d, 1H, H_(arom), J=8.6 Hz), 8.21 (d, 1H, H_(arom), J=5.4 Hz), 8.76 (s,1H, NH_(urea)), 9.12 (s, 1H, NH_(urea)), 12.65 (s, 1H, NH_(lactame)).¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 20.51 (CH₃), 20.93 (CH₃), 30.11(tert-Bu), 31.90 (tert-Bu), 95.00, 108.48, 116.88, 118.10, 118.75,121.77, 122.13, 124.26 (2*C), 126.26, 126.61, 126.67, 127.61, 129.61(2*C), 132.05, 136.04, 136.71, 137.09, 143.72, 144.88, 152.08, 154.51,160.46, 164.11. HRMS (EI): m/z [M+H] calcd for C₃₃H₃₁N₇O₃: 574.2561.Found: 574.2560.

Synthesis 1131-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(3-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-0311

Method F2 was used with 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-2(1H)-oneand 3-tert-butyl-5-isocyanato-1-p-tolyl-1H-pyrazole to afford the titlecompound as a slightly yellow solid (9 mg, 13%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.26 (s, 9H, tert-Bu), 2.37 (s, 3H,Me), 6.32 (s, 1H, H_(arom),), 6.85 (dd, 1H, H_(arom), J=8.1 Hz, J=1.7Hz), 6.89 (d, 1H, H_(Py),5, J=5.3 Hz), 7.19 (d, 1H, H_(arom), J=8.2 Hz),7.31-7.33 (m, 2H, H_(arom)), 7.37-7.40 (m, 3H, H_(arom)), 7.47 (s, 1H,H_(arom),), 8.37-8.41 (m, 3H, H_(arom)), 9.23 (s, 1H, NH_(urea)), 12.54(s, 1H, NH_(lactame)). ¹³C-NMR (δ, ppm, DMSO-d₆): 20.55 (CH₃), 30.16(tert-Bu), 31.95 (tert-Bu), 95.15, 109.72, 110.44, 113.64, 115.03,124.32 (2*C), 129.63 (2*C), 130.46, 135.97, 136.78, 136.82, 141.30,145.25, 151.41, 154.18, 154.65, 160.48. HRMS (EI): m/z [M+H] calcd forC₂₈H₂₇N₇O₃: 510.2248. Found: 510.2250.

Synthesis 1141-(4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-015)

Method F2 was used with2-amino-8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-3(4H)-one and1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound as a slightly pink solid (73 mg, 89%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.32 (d, 1H, H_(arom), J=5.5 Hz),7.27 (d, 1H, H_(arom), J=8.3 Hz), 7.40-7.42 (m, 1H, H_(arom)), 7.54 (t,1H, H_(arom), J=9.8 Hz), 7.59 (t, 1H, H_(arom), J=7.6 Hz), 7.70 (t, 1H,H_(arom), J=8.1 Hz), 7.90 (d, 1H, H_(arom), J=5.5 Hz), 7.93 (d, 1H,H_(arom), J=8.3 Hz), 8.01 (d, 1H, H_(arom), J=8.3 Hz), 8.22 (d, 1H,H_(arom), J=8.6 Hz), 8.69 (dd, 1H, H_(arom), J=7.3 Hz, J=1.9 Hz), 9.27(s, 1H, NH_(urea)), 9.36 (s, 1H, NH_(urea)), 12.59 (s, 1H,NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 106.41, 115.99,116.14, 116.44, 118.47, 119.10, 119.36, 121.67, 121.90, 122.72, 124.89,125.32, 126.38, 126.67, 127.61, 128.74, 130.87, 142.97, 143.57, 146.16,151.73, 152.35, 152.68, 154.32, 157.17. HRMS (EI): m/z [M+H] calcd forC₂₅H₁₆F₄N₆O₃: 525.1293. Found: 525.1292.

Synthesis 1151-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-007)

Method F2 was used with 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-3(4H)-oneand 1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound as a white solid (30 mg, 42%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.61 (d, 1H, H_(Py), J=5.6 Hz), 6.89(dd, 1H, H_(arom), J=8.0 Hz, J=1.9 Hz), 7.26 (d, 1H, H_(arom), J=8.1Hz), 7.39-7.53 (m, 4H, H_(arom)), 8.18 (s, 1H, H_(arom),), 8.37 (d, 1H,H_(Py), J=5.6 Hz), 8.55 (d, 1H, H_(arom), J=7.2 Hz), 8.99 (s, 1H,NH_(urea)), 9.44 (s, 1H, NH_(urea)), 12.94 (s, 1H, NH_(lactame)).¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 106.41, 109.96, 113.96, 115.31,116.00, 116.96, 118.50, 119.56, 122.74, 125.30, 128.43, 130.63, 141.05,145.76, 151.01, 152.06, 152.68, 154.43, 154.66, 156.71, 160.53. HRMS(EI): m/z [M+H] calcd for C₂₁H₁₃F₄N₅O₃: 460.1027. Found: 460.1023.

Synthesis 1161-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(3-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-032)

Method F2 was used with 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-2(1H)-oneand 1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound as a white solid (49 mg, 73%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.87-6.88 (m, 2H, H_(arom)), 7.26 (d,1H, H_(arom), J=7.9 Hz), 7.40-7.54 (m, 4H, H_(arom)), 8.35 (d, 1H,H_(Py),6, J=5.3 Hz), 8.40 (s, 1H, H_(arom),), 8.54 (d, 1H, H_(arom),J=7.2 Hz), 9.05 (s, 1H, NH_(urea)), 9.52 (s, 1H, NH_(urea)), 12.62 (s,1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 109.88, 110.21,113.78, 115.14, 115.92, 116.09, 116.85, 119.46, 122.66, 125.11, 128.36,130.41, 140.91, 144.46, 145.05, 151.96, 152.32, 152.60, 154.22, 154.57,155.06. HRMS (EI): m/z [M+H] calcd for C₂₁H₁₃F₄N₅O₃: 460.1027. Found:460.1025.

Synthesis 1171-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenyl)urea(AA-060)

Method F2 was used with8-(4-amino-3-(methylthio)phenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-oneand 1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene, to afford thetitle compound was obtained as a white solid (5 mg, 12%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): δ 2.50 (s, 3H, CH₃), 2.52 (s, 3H,CH₃), 6.66 (d, 1H, H_(Py), J=5.6 Hz), 7.17 (m, 1H, H_(arom)), 7.35 (d,1H, H_(arom), J=2.7 Hz), 7.42 (m, 2H, H_(arom)), 8.17 (d, 1H, H_(arom),J=8.8 Hz), 8.31 (m, 2H, H_(Py), +H_(arom)), 8.83 (m, 1H, H_(arom)), 9.01(m, 1H, H_(arom)), 11.59 (bs, 1H, NH). LC-MS (m/z): 520 (M+H, 100),rt=2.73 min.

Synthesis 1181-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenyl)urea(AA-061)

Method F2 was used with8-(4-amino-3-(methylthio)phenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-oneand 3-tert-butyl-5-isocyanato-1-tolyl-1H-pyrazole, to afford the titlecompound (9 mg, 20%) as a white solid.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.29 (s, 9H, tert-Bu), 2.35 (s, 3H,CH₃), 2.37 (s, 3H, CH₃), 2.43 (s, 3H, CH₃), 6.44 (s, 1H, CH), 6.56 (d,1H, H_(Py) J=5.6 Hz), 7.06 (dd, 1H, H_(arom), J=8.8 Hz and J=2.7 Hz),7.23-7.27 (m, 3H, H_(arom), J=2.7 Hz), 7.42 (m, 2H, H_(arom)), 7.97 (s,1H, H_(arom)), 8.12 (d, 1H, H_(arom), J=8.8 Hz), 8.22 (d, 1H, H_(Py)J=5.6 Hz), 8.32 (m, 1H, H_(arom)), 11.28 (bs, 1H, NH). LC-MS (m/z): 570(M+H, 100), rt=2.70 min. HRMS (EI): m/z (M+H, 100) calcd forC30H31N7O3S: 570.2281. Found: 570.2282.

Synthesis 1191-(3-tert-butyl-1-(6-methoxypyridin-3-yl)-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-062)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one and5-(3-tert-butyl-5-isocyanato-1H-pyrazol-1-yl)-2-methoxypyridine toafford the title compound (6 mg, 7%) as a white solid.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.77 (s, 9H, tert-Bu), 4.40 (s, 3H,CH₃), 6.93 (s, 1H, CH), 7.15 (d, 1H, H_(Py) J=5.6 Hz), 7.36 (d, 1H,H_(arom), J=8.8 Hz), 7.51 (d, 1H, H_(arom), J=8.4 Hz), 7.59 (dd, 1H,H_(arom), J=11.7 Hz and J=2.6 Hz), 8.31 (dd, 1H, H_(arom), J=8.7 Hz andJ=2.6 Hz), 8.58 (s, 1H, H_(arom)), 8.68 (bs, 1H, H_(arom)), 8.75-8.82(m, 4H, H_(Py)+H_(arom)), 11.95 (bs, 1H, NH). ¹³C-NMR (DMSO-d6), δ(ppm), J (Hz): 40.3, 42.6, 63.6, 106.0, 117.3, 118.5, 118.7, 121.4,126.9, 129.4, 132.5, 132.6, 135.7, 140.5, 146.6, 147.8, 153.3, 156.2,161.5, 161.7, 162.6, 166.5, 171.8, 172.2, 173.6. ¹⁹F-NMR (δ, ppm,DMSO-d₆): −126.99. LC-MS (m/z): 545 (M+H, 100), rt=2.58 min.

Synthesis 1201-(3-tert-butyl-1-(6-methoxypyridin-3-yl)-1H-pyrazol-5-yl)-3-(2-(methylthio)-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-063)

Method F2 was used with8-(4-amino-3-(methylthio)phenoxy)pyrido[3,2-b]pyrazin-3(4H)-one and5-(3-tert-butyl-5-isocyanato-1H-pyrazol-1-yl)-2-methoxypyridine toafford the title compound (97 mg, 53%) as a white powder.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.27 (s, 9H, tert-Bu), 2.43 (s, 3H,CH₃), 3.92 (s, 3H, SCH₃), 6.37 (s, 1H, CH), 6.59 (d, 1H, H_(Py) J=5.6Hz), 6.99 (d, 1H, H_(arom), J=8.8 Hz), 7.03 (dd, 1H, H_(arom), J=8.8 Hz,J=2.6 Hz), 7.21 (d, 1H, H_(arom), J=2.6 Hz), 7.74 (d, 1H, H_(arom),J=8.8 Hz), 7.85 (dd, 1H, H_(arom), J=8.8 Hz, J=2.6 Hz), 8.18 (s, 1H,NH), 8.33 (d, 1H, H_(arom), J=2.6 Hz), 8.35 (d, 1H, H_(Py) J=5.6 Hz),8.37 (s, 1H, CH), 8.98 (s, 1H, NH), 12.94 (s, 1H, NH). ¹³C-NMR(DMSO-d6), δ (ppm), J (Hz): 15.3, 30.0, 31.9, 53.5, 95.3, 106.1, 110.8,117.7, 118.2, 119.3, 124.4, 129.6, 132.0, 133.6, 136.3, 136.6, 142.6,145.3, 149.9, 150.9, 151.9, 152.0, 156.3, 160.7, 161.0, 162.4. LC-MS(m/z): 573 (M+H, 100), rt=2.56 min.

Synthesis 1211-(3-tert-butyl-1-(6-methoxypyridin-3-yl)-1H-pyrazol-5-yl)-3-(4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenyl)urea(AA-064)

Method F2 was used with8-(4-amino-3-(methylthio)phenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-oneand 5-(3-tert-butyl-5-isocyanato-1H-pyrazol-1-yl)-2-methoxypyridine toafford the title compound (33 mg, 35%) as a white powder.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.27 (s, 9H, tert-Bu), 2.43 (s, 3H,CH₃), 2.44 (s, 3H, CH₃), 3.92 (s, 3H, SCH₃), 6.37 (s, 1H, CH), 6.53 (d,1H, H_(Py) J=5.6 Hz), 7.99 (d, 1H, H_(arom), J=8.8 Hz), 7.03 (dd, 1H,H_(arom), J=8.8 Hz, J=2.6 Hz), 7.21 (d, 1H, H_(arom), J=2.6 Hz), 7.76(d, 1H, H_(arom), J=8.8 Hz), 7.85 (dd, 1H, H_(arom), J=8.8 Hz, J=2.6Hz), 8.26 (d, 1H, H_(Py), J=5.6 Hz), 8.33 (d, 1H, H_(arom), J=2.6 Hz),8.35 (s, 1H, NH), 8.96 (s, 1H, NH), 12.76 (s, 1H, NH). ¹³C-NMR(DMSO-d6), δ (ppm), J (Hz): 15.3, 20.3, 30.0, 31.9, 53.5, 95.3, 105.8,110.7, 117.5, 117.8, 119.4, 124.3, 129.5, 131.9, 133.5, 136.3, 137.6,142.6, 145.5, 149.9, 150.4, 151.8, 156.2, 158.8, 159.9, 161.0, 162.3.LC-MS (m/z): 587 (M+H, 100), rt=2.63 min. HRMS (EI): m/z (M+H, 100)calcd for C29H30N8O4S: 587.2183. Found: 587.2186.

Synthesis 1221-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(4-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenyl)urea(AA-065)

Method F2 was used with8-(4-amino-3-(methylthio)phenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-oneand 3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound (45 mg, 51%) as a white solid.

¹H-NMR (acetone-d6), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 2.43 (s,3H, CH₃), 2.44 (s, 3H, CH₃), 6.37 (s, 1H, CH), 6.54 (d, 1H, H_(Py) J=5.6Hz), 7.02 (dd, 1H, H_(arom), J=8.8 Hz and J=2.6 Hz), 7.21 (d, 1H,H_(arom), J=2.6 Hz), 7.39-7.42 (m, 1H, H_(arom)), 7.53-7.55 (m, 4H,H_(arom)), 7.77 (d, 1H, H_(arom), J=8.8 Hz), 8.27 (d, 1H, H_(Py) J=5.6Hz), 8.37 (s, 1H, NH), 8.98 (s, 1H, NH), 12.75 (bs, 1H, NH). ¹³C-NMR(DMSO-d6), δ (ppm), J (Hz): 15.3, 20.3, 30.0, 31.9, 96.2, 105.8, 117.5,117.8, 119.5, 123.9 (2), 124.2, 127.0, 129.1 (2), 131.8, 133.6, 136.8,138.6, 145.5, 149.8, 150.4, 152.0, 156.2, 158.8, 159.9, 160.7. LC-MS(m/z): 556 (M+H, 100), rt=2.66 min. HRMS (EI): m/z (M+H, 100) calcd forC29H29N7O3S: 556.2125. Found: 556.2125.

Synthesis 1231-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-(methylthio)-4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-066)

Method F2 was used with8-(4-amino-3-(methylthio)phenoxy)pyrido[2,3-b]pyrazin-2(1H)-one and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound (54 mg, 59%) was obtained as a pale brown powder.

¹H-NMR (CDCl₃), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 2.43 (s, 3H,CH₃), 6.36 (s, 1H), 6.88 (d, 1H, H_(Py) J=5.3 Hz), 7.06 (dd, 1H,H_(arom), J=8.8 Hz, J=2.7 Hz), 7.24 (d, 1H, H_(arom), J=2.7 Hz),7.39-7.43 (m, 1H, H_(arom)), 7.52-7.55 (m, 4H, H_(arom)), 7.78 (d, 1H,H_(arom), J=8.8 Hz), 8.36 (m, 1H, H_(arom)), 8.38 (s, 1H, NH or CH),8.41 (s, 1H, NH or CH), 8.98 (s, 1H, NH), 12.55 (bs, 1H, NH). ¹³C-NMR(DMSO-d6), δ (ppm), J (Hz): 15.5, 30.0, 31.9, 96.2, 110.0, 117.8, 119.6,123.9 (3), 124.1, 127.0, 129.1 (3), 131.6, 133.8, 136.8, 138.5, 144.0,145.2, 149.7, 152.0, 154.4, 156.1, 160.7. LC-MS (m/z): 542 (M+H, 100),rt=2.52 min. HRMS (EI): m/z (M+H, 100) calcd for C28H27N7O3S: 542.1968.Found: 542.1969.

Synthesis 1241-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-(methylthio)-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-067)

Method F2 was used with8-(4-amino-3-(methylthio)phenoxy)pyrido[3,2-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound (175 mg, 97%) as a white powder.

¹H-NMR (CDCl₃-d6), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 2.43 (s, 3H,CH₃), 6.36 (s, 1H), 6.60 (d, 1H, H_(Py,5), J=5.6 Hz), 7.03 (dd, 1H,H_(arom), J=8.8 Hz, J=2.7 Hz), 7.21 (d, 1H, H_(arom), J=2.7 Hz),7.39-7.43 (m, 1H, H_(arom)), 7.53-7.54 (m, 4H, H_(arom)), 7.77 (d, 1H,H_(arom), J=8.8 Hz), 8.18 (s, 1H, NH or CH), 8.35 (d, 1H, H_(Py) J=5.6Hz), 8.37 (s, 1H, NH or CH), 8.98 (s, 1H, NH or CH), 12.89 (s, 1H, NH).¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 15.3, 30.0 (3), 31.9, 96.2, 106.1,117.7, 118.2, 119.3, 123.9 (2), 124.3, 127.0, 129.1 (2), 131.8, 133.7,136.8, 138.5, 145.3, 149.9, 150.8, 152.0 (2), 156.3, 160.6, 160.7. LC-MS(m/z): 542 (M+H, 100), rt=2.60 min. HRMS (EI): m/z (M+H, 100) calcd forC28H27N7O3S: 542.1968. Found: 542.1968.

Synthesis 1251-(4-(3-(dimethylamino)pyrido[3,2-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-068)

Method F2 was used with8-(4-amino-3-fluorophenoxy)-N,N-dimethylpyrido[3,2-b]pyrazin-3-amine and1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound as a white solid. Yield: 40 mg (66%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 3.28 (s, 6H, N(CH₃)₂), 6.65 (d,J=5.3, 1H, H_(Py)), 7.05 (m, 2H, H_(arom)), 7.30 (m, 1H, H_(arom)), 7.41(m, 1H, H_(arom)), 7.51 (m, 1H, H_(arom)), 8.22 (m, 1H, H_(arom)), 8.60(d, J=5.3, 1H, H_(Py)), 8.65 (m, 1H, H_(arom)), 8.71 (s, 1H, H_(arom)),9.19 (s, 1H, NH), 9.36 (s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz):27.1, 105.3, 108.3 (d, J_(FC)=22.3), 116.1 (d, J_(FC)=20.5), 116.3 (d,J_(FC)=2.6), 116.6 (m), 119.4 (m), 122.0 (d, J_(FC)=2.3), 122.8, 125.0,125.4 (m), 128.5 (d, J_(FC)=11.4), 139.6 (br), 149.6 (d, J_(FC)=10.3),152.0, 152.4 (d, H_(FC)=245), 152.8, 153.4, 153.4 (d, J_(FC)=248),155.3, 160.4; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −60.8, −124.0, −125.3; LC-MS(2.28 min): m/z 505.2 (M+H, 100); HRMS (2.80 min): m/z calcd. forC₂₃H₁₇F₆N₆O₂ [M+H⁺]: 505.14059. Found: 505.13996.

Synthesis 1261-(3-Tea-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(4-(3-(dimethylamino)pyrido[3,2-b]pyrazin-8-yloxy)-2-fluorophenyl)urea(AA-070)

Method F2 was used with8-(4-amino-3-fluorophenoxy)-N,N-dimethylpyrido[3,2-b]pyrazin-3-amine and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to give the product as alight yellow solid. Yield: 65 mg (90%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.29 (s, 9H, tert-Bu), 3.28 (s, 6H,N(CH₃)₂), 6.41 (s, 1H, H_(arom)), 6.62 (d, 1H, J=5.2, H_(Py)), 7.02 (m,1H, H_(arom)), 7.26 (m, 1H, H_(arom)), 7.44 (m, 1H, H_(arom)), 7.55 (m,4H, H_(arom)), 8.14 (m, 1H, H_(arom)), 8.59 (d, 1H, J=5.2, H_(Py)), 8.71(s, 1H, H_(arom)), 8.87 (s, 1H, NH), 8.99 (s, 1H, NH); ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 30.2, 32.0, 37.4, 95.2, 105.3, 108.3 (d,J_(FC)=22.3), 116.2, 121.8, 121.9, 124.4 (d, J_(FC)=10.7), 124.5, 127.4,129.3, 136.0, 137.0, 138.5, 149.4 (d, J_(FC)=10.2), 151.4, 152.4, 152.4(d, J_(FC)=245), 153.5, 154.2, 160.4, 160.8; ¹⁹F-NMR (DMSO-d₆), δ (ppm):−125.3; LC-MS (2.25 min): m/z 541.1 (M+H, 100); HRMS (2.85 min): m/zcalcd. for C₂₉H₂₉FN₈NaO₂ [M+Na⁺]: 563.22897. Found: 563.22865.

Synthesis 1273-tert-butyl-1-(4-(methylsulfonyl)phenyl)-1H-pyrazol-5-amine

4-(methylsulfonyl)phenylhydrazine hydrochloride (1.133 g, 5.09 mmol) and4,4-dimethyl-3-oxopentanenitrile (0.697 g, 5.57 mmol) were weighed intoa 100 mL RBF. 0.2 M HCl in EtOH (42 mL) was added and the suspension washeated to reflux for 27 h, during which time all solids graduallydissolved to give a yellow solution. The solution was diluted with 1 MNaOH_((aq)) (˜16 mL) to pH 12-13, EtOAc (70 mL) was added and thebiphasic system was vigorously stirred for 5 min. The organic layer wasisolated, dried (MgSO₄), filtered and concentrated to give a yellowcrystalline solid. Yield: 1.42 g (95%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.23 (s, 9H, tert-Bu), 3.22 (s, 3H,Me), 5.45 (br s, 2H, NH₂), 5.46 (s, 1H, H_(Pyz)), 7.90 (d, 2H, J=8.7,H_(arom)), 7.98 (d, 2H, J=8.7, H_(arom));

¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 30.0, 31.9, 43.7, 88.3, 121.6,128.1, 136.7, 143.8, 148.0, 162.2; LC-MS (1.98 min): m/z 294.1 (M+H,100).

Synthesis 128 Solution of3-tert-butyl-5-isocyanato-1-(4-(methylsulfonyl)phenyl)-1H-pyrazole inCH₂Cl₂

3-tert-butyl-1-(4-(methylsulfonyl)phenyl)-1H-pyrazol-5-amine (295 mg,1.01 mmol) was weighed into a 100 mL RBF and CH₂Cl₂ (20 mL) andsaturated aqueous NaHCO₃ (20 mL) were added. The resulting biphasicsystem was stirred and cooled to 0° C., and subsequently treateddropwise with 1.9 M phosgene in toluene (1.06 mL, 2.02 mmol) over 30 s.The mixture was stirred vigorously for 10 min, the organic phase wasisolated, washed with H₂O (20 mL), dried (MgSO₄), filtered andconcentrated to 10 mL to give a 100 mM solution of the title compound.IR (v, cm⁻¹): 2260 (N═C═O).

Synthesis 1291-(3-tert-butyl-1-(4-(methylsulfonyl)phenyl)-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-090)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one (56 mg, 0.206mmol) and a 0.1 M solution of3-tert-butyl-5-isocyanato-1-(4-(methylsulfonyl)phenyl)-1H-pyrazole inCH₂Cl₂ (5.8 mL, 0.58 mmol). The title compound was obtained as a yellowsolid in 41% yield (50 mg) after chromatography on a Biotage 25+Mcolumn.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.30 (s, 9H, tert-Bu), 3.27 (s, 3H,SO₂CH₃), 6.46 (s, 1H, H_(Py)), 6.65 (d, J=5.6, 1H, H_(Pyz)), 7.05 (m,1H, H_(arom)), 7.30 (m, 1H, H_(arom)), 7.85 (d, J=8.7, 2H, H_(arom)),8.08 (d, J=8.7, 2H, H_(arom)), 8.12 (m, 1H, H_(arom)), 8.17 (s, 1H,H_(arom)), 8.37 (d, J=5.6, 1H, H_(Py)), 8.97 (s, 1H, NH), 8.99 (s, 1H,NH), 12.90 (s, 1H, NH);

¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 30.0, 32.1, 43.5, 97.0, 106.5, 108.5(d, J_(FC)=22.4), 116.4, 118.4, 122.0, 123.9, 124.7 (d, J_(FC)=10.8),128.3, 137.4, 138.7, 142.5, 145.6, 148.8 (d, J_(FC)=10.5), 151.1, 151.5,152.2, 152.5 (d, J_(FC)=245), 156.6, 160.5, 162.1; ¹⁹F-NMR (DMSO-d₆), δ(ppm): −124.3; LC-MS (m/z): LC-MS 592.1 (M+H, 100), rt=2.44 min; HRMS(7.17 min): m/z calcd. for C₂₈H₂₇FN₇O₆S (M+H, 100)⁺: 461.09798. Found:461.09771.

Synthesis 1301-(3-tert-butyl-1-(4-(methylsulfonyl)phenyl)-1H-pyrazol-5-yl)-3-(4-(2,3-dioxo-1,2,3,4-tetrahydropyrido[3,2-b]pyrazin-8-yloxy)-2-fluorophenyl)urea(AA-092)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[3,2-b]pyrazine-2,3(1H,4H)-dione (58mg, 101 μmol) and a 0.06 M solution of3-tert-butyl-5-isocyanato-1-(4-(methylsulfonyl)phenyl)-1H-pyrazole inCH₂Cl₂ (6.8 mL, 0.41 mmol). The title compound was obtained as a whitesolid. Yield: 30 mg (49%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.30 (s, 9H, tert-Bu), 3.27 (s, 3H,SO₂CH₃), 6.45 (s, 1H, H_(Py)), 6.57 (d, 1H, J=5.7, H_(Py)), 7.00 (m, 1H,H_(arom)), 7.22 (m, 1H, H_(arom)), 7.85 (d, 2H, J=8.7, H_(arom)), 7.95(d, 1H, J=5.7, H_(Py)), 8.07 (d, 2H, J=8.7, H_(arom)), 8.09 (m, 1H,H_(arom)), 8.94 (s, 1H, NH), 8.97 (s, 1H, H_(arom)), 11.89 (s, 1H, NH),12.38 (s, 1H, NH); ¹⁹F-NMR (DMSO-d₆), δ (ppm): −124.6; LC-MS (m/z):608.1 (M+H, 100), rt=2.39 min; HRMS (3.07 min): m/z calcd. forC₂₈H₂₇FN₇O₆S [M+H⁺]: 608.17221. Found: 608.17142.

Synthesis 1311-(4-(2,3-Dioxo-1,2,3,4-tetrahydropyrido[3,2-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-072)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[3,2-b]pyrazine-2,3(1H,4H)-dione and1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to give the titlecompound as a beige solid. Yield: 52 mg (51%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.60 (d, J=5.4, 1H, H_(Py)), 7.04 (m,2H, H_(arom)), 7.27 (m, 1H, H_(arom)), 7.41 (m, 1H, H_(arom)), 7.52 (m,1H, H_(arom)), 7.98 (d, J=5.4, 1H, H_(Py)), 8.22 (m, 1H, H_(arom)), 8.64(m, 1H, H_(arom)), 9.19 (s, 1H, NH), 9.35 (s, 1H, NH), 11.92 (s, 1H,NH), 12.40 (s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 106.6, 108.2(d, J_(FC)=22.5), 112.5, 116.0, 116.2, 116.6 (M), 119.5 (m), 120.6,121.9 (d, J_(FC)=2.3), 123.9 (qua, J_(FC)=270), 124.3 (d, J_(FC)=10.6),125.4 (m), 128.5 (d, J_(FC)=11.4), 140.6, 143.2, 149.0 (d, J_(FC)=10.4),150.2, 152.0, 152.3 (d, J_(FC)=245), 153.4 (d, J_(FC)=249), 154.7,156.0; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −60.2, −123.5, −124.9; LC-MS (m/z):494.0 (M+H, 100), rt=2.57 min; HRMS (3.06 min): m/z calcd. forC₂₁H₁₂F₅N₅NaO₄ [M+Na⁺]: 516.07017. Found: 516.06998.

Synthesis 1321-(3-Tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-morpholinopyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-071)

A solution of2-fluoro-4-(3-morpholinopyrido[2,3-b]pyrazin-8-yloxy)aniline (47 mg, 138μmol) in dry THF (5 mL) was treated with a solution of3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole (1.1 mL of a 0.25 Msolution in CH₂Cl₂, 275 μmol) at 0° C. A yellow precipitate started toform gradually and after 1 h at RT, hexane (20 mL) was added and theyellow precipitate was filtered off. It was redissolved in MeOH/CH₂Cl₂(1:1), evaporated onto silica gel and the product was eluted with agradient of 0% to 20% MeOH in EtOAc to give the product as a lightyellow solid.

Yield: 71 mg (89%).

1H-NMR (DMSO-d6), (ppm), J (Hz): 1.30 (s, 9H, tert-Bu), 3.77 (m, 4H,N(CH₂CH₂)₂O), 3.84 (m, 4H, N(CH₂CH₂)₂O), 6.41 (s, 1H, H_(arom)), 6.68(d, 1H, J=5.2 Hz, H_(Py)), 7.02 (m, 1H, H_(arom)), 7.27 (m, 1H,H_(arom)), 7.44 (m, 1H, H_(arom)), 7.55 (m, 4H, H_(arom)), 8.15 (m, 1H,H_(arom)), 8.63 (d, 1H, J=5.2, H_(arom)), 8.84 (m, 2H,H_(arom)+H_(urea)), 8.98 (s, 1H, H_(urea)); ¹³C-NMR (DMSO-d6), (ppm), J(Hz): 30.2, 32.0, 44.4, 65.9, 95.1, 105.9, 108.3 (d, J_(FH)=22.3),116.3, 121.8, 122.5, 124.4 (d, J_(FH)=10.7), 124.5, 127.4, 129.3, 136.3,137.0, 138.3, 149.3 (d, J_(FH)=10.4), 151.4, 152.1, 152.4 (d,J_(FH)=245), 153.7, 153.9, 160.4, 160.8; ¹⁹F-NMR (DMSO-d6), (ppm):−125.3; LC-MS (m/z): 583.1 (M+H, 100), rt=2.33 min; HRMS (2.88 min): m/zcalcd. for C31H32FN8O3 [M+H+]: 583.25759. Found: 583.25719;

Synthesis 1331-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-morpholinopyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-073)

Method F2 was used with2-fluoro-4-(3-morpholinopyrido[2,3-b]pyrazin-8-yloxy)aniline and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to give the product as alight yellow solid. Yield: 97 mg (90%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.29 (s, 9H, tert-Bu), 2.39 (s, 3H,CH₃), 3.77 (m, 4H, N(CH₂CH₂)₂O), 3.84 (m, 4H, N(CH₂CH₂)₂O), 6.40 (s, 1H,H_(Pyz)), 6.67 (d, 1H, J=5.3, _(Py)rH), 7.03 (m, 1H, H_(arom)), 7.27 (m,1H, H_(arom)), 7.35 (d, 2H, J=8.3, H_(arom)), 7.41 (d, 2H, J=8.3,H_(arom)), 8.17 (m, 1H, H_(arom)), 8.62 (d, 1H, J=5.3, H_(Py)), 8.80 (s,1H, NH), 8.84 (s, 1H, H_(arom)), 9.00 (s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ(ppm), J (Hz): 20.6, 30.2, 32.0, 44.4, 65.9, 94.6, 105.8, 108.3 (d,J_(FC)=22.4), 116.2, 121.7, 122.5, 124.5, 124.6 (d, J_(FC)=10.7), 129.7,135.9, 136.3, 136.9 (d, J_(FC)=5.7), 149.2 (d, J_(FC)=10.4), 151.3,152.1, 152.3 (d, J_(FC)=245), 153.7, 153.9, 160.4, 160.8; ¹⁹F-NMR(DMSO-d₆), δ (ppm): −124.8; LC-MS (m/z): 597.2 (M+H, 100), rt=2.43 min;HRMS (3.01 min): m/z calcd. for C₃₂H₃₄FN₈O₃ [M+H⁺]: 597.27324. Found:597.27289.

Synthesis 1341-(3-Tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(2,3-dioxo-1,2,3,4-tetrahydropyrido[3,2-b]pyrazin-8-yloxy)-2-fluorophenyl)urea(AA-074)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[3,2-b]pyrazine-2,3(1H,4H)-dione and3-tert-butyl-5-isocyanato-1-p-tolyl-1H-pyrazole to give the titlecompound as a white solid. Yield: 33 mg (44%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 2.36 (s, 3H,CH₃), 6.37 (s, 1H, _(Py)razoleH), 6.53 (d, 1H, J=5.3, H_(Py)), 6.91 (m,1H, H_(arom)), 7.08 (m, 1H, H_(arom)), 7.29 (d, 2H, J=8.3, H_(arom)),7.38 (d, 2H, J=8.3, H_(arom)), 7.90 (d, 1H, J=5.3, H_(Py)), 8.07 (m, 1H,H_(arom)), 9.14 (br s, 1H, NH), 9.24 (br s, 1H, H_(arom)), 12.00 (br s,2H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 20.6, 30.2, 32.0, 94.8,106.6, 107.8 (d, J_(FC)=22.4), 115.8, 122.2, 124.3 (d, J_(FC)=10.7),129.5, 136.0, 136.7, 137.2, 141.1, 142.3 (br), 149.2 (d, J_(FC)=9.8),150.7, 151.5, 151.6, 152.3 (d, J_(FC)=245), 153.5, 156.3 (br), 156.4,160.5; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −124.4; LC-MS (m/z): 544.0 (M+H,100), rt=2.62 min; HRMS (3.01 min): m/z calcd. for C₂₈H₂₇FN₇O₄ [M+H⁺]:544.21031. Found: 544.21063.

Synthesis 1351-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-075)

Method F2 was used with 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-3(4H)-oneand 3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound as a slightly yellow solid (97 mg, 62%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.27 (s, 9H, tert-Bu), 6.35 (s, 1H,H_(arom),), 6.61 (d, 1H, H_(Py), J=5.6 Hz), 6.84 (dd, 1H, H_(arom),J=1.8 Hz, J=8.0 Hz), 7.20 (d, 1H, H_(arom), J=1.2 Hz, J=8.1 Hz),7.36-7.42 (m, 2H, H_(arom)), 7.44 (t, 1H, H_(arom), J=2.1 Hz), 7.52-7.53(m, 4H, H_(arom)), 8.18 (s, 1H, H_(arom),), 8.36 (d, 1H, H_(Py), J=5.6Hz), 8.44 (s, 1H, NH_(urea)), 9.23 (s, 1H, NH_(urea)), 12.89 (s, 1H,NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 30.04 (tert-Bu),31.89 (tert-Bu), 95.63, 106.54, 109.58, 113.54, 114.96, 118.38, 124.14(2*C), 127.13, 129.14 (2*C), 130.46, 136.76, 138.40, 141.25, 145.42,151.05, 151.42, 152.01, 154.34, 156.35, 160.40, 160.65.

Synthesis 1361-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(3-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-076)

Method F2 was used with 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-2(1H)-oneand 3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound as a yellow solid (57 mg, 29%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.27 (s, 9H, tert-Bu), 6.35 (s, 1H,H_(arom),), 6.85 (dd, 1H, H_(arom), J=8.1 Hz, J=1.9 Hz), 6.88 (d, 1H,H_(Py), J=5.3 Hz), 7.20 (dd, 1H, H_(arom), J=8.2 Hz, J=1.2 Hz),7.37-7.42 (m, 2H, H_(arom)), 7.47 (t, 1H, H_(arom), J=2.0 Hz), 7.52-7.53(m, 4H, H_(arom)), 8.35-8.43 (m, 3H, H_(arom)), 9.23 (s, 1H, NH_(urea)),12.54 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 30.04(tert-Bu), 31.90 (tert-Bu), 105.11, 106.87, 109.67, 110.38, 111.17,113.58, 115.00, 119.52, 124.14 (2*C), 127.35, 129.14 (2*C), 130.38,136.77, 138.41, 141.21, 145.22, 151.42, 154.09, 154.55, 155.88, 160.66.

Synthesis 1371-(4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea(AA-077)

Method F2 was used with2-amino-8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-tolyl-1H-pyrazole to afford the titlecompound as a beige solid (46 mg, 51%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 2.40 (s, 3H,Me), 6.30 (d, 1H, H_(arom), J=5.5 Hz), 6.39 (s, 1H, H_(arom)), 7.22 (d,1H, H_(arom), J=8.3 Hz), 7.37 (d, 2H, H_(arom), J=8.2 Hz), 7.46 (d, 2H,H_(arom), J=8.3 Hz), 7.56 (t, 1H, H_(arom), J=7.5 Hz), 7.63 (t, 1H,H_(arom), J=8.0 Hz), 7.86-7.91 (m, 3H, H_(arom)), 8.06 (d, 1H, H_(arom),J=8.5 Hz), 8.72 (s, 1H, NHurea), 9.05 (s, 1H, NHurea), 12.58 (s, 1H,NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 20.50 (CH₃), 30.11(tert-Bu), 31.90 (tert-Bu), 95.11, 106.44, 116.08, 118.75, 119.35,121.56, 122.26, 124.22 (2*C), 126.33, 126.49, 126.55, 127.85, 129.58(2*C), 131.26, 136.07, 136.67, 137.12, 142.95, 143.56, 146.12, 151.71,152.21, 152.72, 157.14, 160.43. HRMS (EI): m/z [M+H] calcd forC₃₂H₃₀N₈O₃: 575.2514. Found: 575.2519.

Synthesis 1381-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(3-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-078)

Method F2 was used with8-(3-aminophenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-p-tolyl-1H-pyrazole to afford the titlecompound as a white solid (89 mg, 57%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.26 (s, 9H, tert-Bu), 2.36 (s, 3H,Me), 2.42 (s, 3H, Me), 6.32 (s, 1H, H_(arom),), 6.55 (d, 1H, H_(Py),J=6.6 Hz), 6.83 (dd, 1H, H_(arom), J=2.2 Hz, J=8.1 Hz), 7.21 (d, 1H,H_(arom), J=7.9 Hz), 7.31 (d, 2H, H_(arom), J=8.3 Hz), 7.36 (s, 1H,H_(arom)), 7.37 (d, 2H, H_(arom), J=8.3 Hz), 7.44 (t, 1H, H_(arom),J=2.1 Hz), 8.27 (d, 1H, H_(Py), J=6.8 Hz), 8.48 (s, 1H, NH_(urea)), 9.30(s, 1H, NH_(urea)), 12.75 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ(ppm), J (Hz): 20.42 (CH₃), 20.46 (CH₃), 30.08 (tert-Bu), 31.87(tert-Bu), 95.67, 106.19, 109.72, 113.56, 114.91, 117.74, 124.08 (2*C),129.49 (2*C), 130.38, 136.03, 136.51, 136.76, 141.44, 145.71, 150.40,151.61, 154.30, 156.32, 158.98, 159.67, 160.33. HRMS (EI): m/z [M+H]calcd for C₂₉H₂O₇O₃: 524.2405. Found: 524.2409.

Synthesis 1391-(4-(3-amino-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-079)

Method F2 was used with3-amino-8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-2(1H)-one and1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound as a yellow/orange solid (31 mg, 38%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.30 (d, 1H, H_(arom), J=5.5 Hz),7.34 (d, 1H, H_(arom), J=8.3 Hz), 7.41 (s, 1H, H_(arom)), 7.54 (t, 1H,H_(arom), J=8.7 Hz), 7.60 (t, 1H, H_(arom), J=7.4 Hz), 7.71 (t, 1H,H_(arom), J=7.2 Hz), 7.99 (d, 1H, H_(arom), J=8.8 Hz), 8.01 (d, 1H,H_(arom), J=5.6 Hz), 8.04 (d, 1H, H_(arom), J=8.3 Hz), 8.24 (d, 1H,H_(arom), J=8.6 Hz), 8.69 (d, 1H, H_(arom), J=7.2 Hz), 9.32 (s, 1H,NH_(urea)), 9.38 (s, 1H, NH_(urea)), 12.39 (s, 1H, NH_(lactame)).¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 104.61, 113.94, 116.00, 116.47,118.03, 119.16, 121.96, 122.73, 124.89, 125.33, 126.53, 126.72, 127.44,128.73, 131.24, 144.29, 145.44, 146.76, 151.08, 152.38, 152.60, 154.35,154.77. HRMS (EI): m/z [M+H] calcd for C25H16F4N6O3: 525.1293. Found:525.1292.

Synthesis 1401-(4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)-3-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)urea(AA-080)

Method F2 was used with2-amino-8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-3(4H)-one and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound as a slightly pink solid (70 mg, 80%). ¹H-NMR (DMSO-d6), δ(ppm), J (Hz): 1.29 (s, 9H, tert-Bu), 6.31 (d, 1H, H_(arom), J=5.5 Hz),6.41 (s, 1H, H_(arom)), 7.22 (d, 1H, H_(arom), J=8.3 Hz), 7.44 (t, 1H,J=7.0 Hz), 7.54-7.65 (m, 6H, H_(arom)), 7.85 (d, 1H, H_(arom), J=8.3Hz), 7.88-7.91 (m, 2H, H_(arom)), 8.06 (d, 1H, H_(arom), J=8.6 Hz), 8.76(s, 1H, NH_(urea)), 9.04 (s, 1H, NH_(urea)), 12.58 (s, 1H,NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 30.08 (tert-Bu),31.92 (tert-Bu), 95.69, 106.46, 116.05, 118.94, 119.36, 121.56, 122.27,124.12 (2*C), 126.32, 126.52, 126.55, 127.11, 127.97, 129.17 (2*C),131.22, 137.15, 138.59, 142.95, 143.56, 146.22, 151.71, 152.33, 152.72,157.12, 160.68. HRMS (EI): m/z [M+H] calcd for C₃₁H₂O₈O₃: 561.2357.Found: 561.2351.

Synthesis 1411-(4-(3-amino-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)-3-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)urea(AA-081)

Method F2 was used with3-amino-8-(4-aminonaphthalen-1-yloxy)pyrido[2,3-b]pyrazin-2(1H)-one and3-tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole to afford the titlecompound as a yellow/orange solid (46 mg, 44%). ¹H-NMR (DMSO-d6), δ(ppm), J (Hz): 1.30 (s, 9H, tert-Bu), 6.28 (d, 1H, H_(arom), J=5.5 Hz),6.42 (s, 1H, H_(arom)), 7.31 (d, 1H, H_(arom), J=8.3 Hz), 7.44 (t, 1H,J=7.1 Hz), 7.55-7.66 (m, 6H, H_(arom)), 7.89-7.96 (m, 2H, H_(arom)),7.99 (d, 1H, H_(arom), J=5.5 Hz), 8.08 (d, 1H, H_(arom), J=8.6 Hz), 8.82(s, 1H, NH_(urea)), 9.11 (s, 1H, NH_(urea)), 12.38 (s, 1H,NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 30.08 (tert-Bu),31.93 (tert-Bu), 95.78, 104.61, 113.91, 116.44, 118.37, 121.92, 122.14,124.09 (2*C), 126.45, 126.47, 126.54, 127.09, 127.73, 129.15 (2*C),131.63, 137.13, 138.59, 144.28, 145.43, 146.74, 151.01, 151.12, 152.33,154.75, 160.69. HRMS (EI): m/z [M+H] calcd for C₃₁H₂₈N₈O₃: 561.2357.Found: 561.2350.

Synthesis 1421-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(4-(3-oxo-2-(trifluoromethyl)-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-082)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)-2-(trifluoromethyl)pyrido[2,3-b]pyrazin-3(4H)-oneand 1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound as a slightly yellow solid (31 mg, 45%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.40 (d, 1H, H_(arom), J=5.7 Hz),7.41-7.42 (m, 1H, H_(arom)), 7.49 (d, 1H, H_(arom), J=8.3 Hz), 7.54 (t,1H, H_(arom), J=9.9 Hz), 7.60 (t, 1H, H_(arom), J=7.6 Hz), 7.73 (t, 1H,H_(arom), J=7.7 Hz), 7.85 (d, 1H, H_(arom), J=8.4 Hz), 8.15 (d, 1H,H_(arom), J=8.3 Hz), 8.29 (d, 1H, H_(arom), J=8.6 Hz), 8.38 (d, 1H,H_(arom), J=5.7 Hz), 8.71 (d, 1H, H_(arom), J=6.0 Hz), 9.39 (s, 1H,NHurea), 9.42 (s, 1H, NHurea), 13.55 (s, 1H, NH_(lactame)). ¹³C-NMR(DMSO-d6), δ (ppm), J (Hz): 105.66, 116.02, 116.54, 117.64, 118.82,119.29, 121.01, 121.40, 122.08, 122.72, 124.88, 125.35, 126.15, 126.86,127.63, 128.65, 132.23, 143.10, 144.37, 146.85, 152.39, 152.48, 153.32,154.36, 154.82, 162.35. HRMS (EI): m/z [M+H] calcd for C₂₆H₁₄F₇N₅O₃:578.1058. Found: 578.1064.

Synthesis 1431-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(4-(2-oxo-3-(trifluoromethyl)-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)naphthalen-1-yl)urea(AA-083)

Method F2 was used with8-(4-aminonaphthalen-1-yloxy)-3-(trifluoromethyl)pyrido[2,3-b]pyrazin-2(1H)-oneand 1-fluoro-2-isocyanato-4-(trifluoromethyl)benzene to afford the titlecompound was obtained as a slightly yellow solid (5 mg, 5%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.79 (d, 1H, H_(arom), J=5.3 Hz),7.42-7.43 (m, 1H, H_(arom)), 7.47 (d, 1H, H_(arom), J=8.3 Hz), 7.55 (t,1H, H_(arom), J=9.8 Hz), 7.62 (t, 1H, H_(arom), J=7.6 Hz), 7.74 (t, 1H,H_(arom), J=7.9 Hz), 7.97 (d, 1H, H_(arom), J=8.5 Hz), 8.14 (d, 1H,H_(arom), J=8.3 Hz), 8.28 (d, 1H, H_(arom), J=8.6 Hz), 8.40 (d, 1H,H_(arom), J=5.2 Hz), 9.71 (dd, 1H, H_(arom), J=1.8, 7.2 Hz), 9.38 (s,1H, NH_(urea)), 9.42 (s, 1H, NH_(urea)), 13.51 (s, 1H, NH_(lactame)).

¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 110.67, 116.02, 116.48, 117.14,117.76, 118.64, 119.21, 120.84, 121.88, 122.73, 123.05, 124.89, 125.40,126.13, 126.87, 127.05, 127.37, 128.67, 131.99, 141.78, 144.58, 146.66,151.64, 152.39, 152.53, 154.36. HRMS (EI): m/z [M+H] calcd forC₂₆H₁₄F₇N₅O₃: 578.1058. Found: 578.1051.

(VII) Synthesis of Amides 1. Amides from Common Intermediates Synthesis144N-(3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)-3-(trifluoro-methoxy)benzamide(AA-002)

Method G1: 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one (43 mg,0.169 mmol) and diisopropylethylamine (44 μL, 0.254 mmol) were mixed indry THF (5.0 mL) and 3-trifluoromethoxybenzoyl chloride (57 mg, 0.254mmol) was added. This mixture was heated to reflux for 17 h. Aftercooling at RT, the solvent was removed in vacuo. The obtained oilyresidue was dissolved in DCM and washed with water and dried over MgSO₄.After evaporation of DCM, the residue was retaken in Et₂O, trituratedand filtered off to afford the title compound as a white solid (45 mg,60%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.68 (d, 1H, H_(Py), J=5.6 Hz), 7.01(ddd, 1H, H_(arom), J=8.1 Hz, J=2.3 Hz, J=0.7 Hz), 7.50 (t, 1H,H_(arom), J=8.2 Hz), 7.62 (d, 1H, H_(arom), J=8.3 Hz), 7.68-7.72 (m, 3H,H_(arom)), 7.89 (5, 1H, H_(arom)), 7.99 (d, 1H, H_(arom), J=7.9 Hz),8.19 (s, 1H, H_(arom),), 8.39 (d, 1H, H_(Py),6, J=5.6 Hz), 10.53 (s, 1H,NH_(amide)), 12.91 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J(Hz): 106.78, 111.77, 115.45, 117.10, 118.48, 120.08, 120.95, 124.07,126.69, 130.39, 130.54, 136.68, 140.54, 145.47, 148.17, 151.16, 152.08,154.21, 156.36, 160.28, 163.97. HRMS (EI): m/z [M+H] calcd forC₂₁H₃F₃N₄O₄: 443.0962. Found: 443.0950.

Synthesis 1453-tert-butyl-N-(3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)benzamide(AA-003)

Method G1 was used with 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-3(4H)-oneand 3-tertbutylbenzoyl chloride to afford the title compound as a whitesolid (29 mg, 45%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.33 (s, 9H, tert-Bu), 6.67 (d, 1H,H_(Py),5, J=5.6 Hz), 6.98 (ddd, 1H, H_(arom), J=8.1 Hz, J=2.4 Hz, J=0.8Hz), 7.46 (t, 1H, H_(arom), J=7.7 Hz), 7.48 (t, 1H, H_(arom), J=8.2 Hz),7.63 (d, 1H, H_(arom), J=7.9 Hz), 7.70 (d, 1H, H_(arom), J=8.2 Hz),7.74-7.77 (m, 2H, H_(arom)), 7.90 (t, 1H, H_(arom), J=1.7 Hz), 8.19 (s,1H, H_(arom),), 8.39 (d, 1H, H_(Py),6, J=5.6 Hz), 10.36 (s, 1H,NH_(amide)), 12.91 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J(Hz): 31.00 (tert-Bu), 34.56 (tert-Bu), 106.78, 111.84, 115.20, 117.18,118.53, 124.30, 124.79, 128.11, 128.67, 130.37, 134.44, 141.05, 145.54,150.95, 151.20, 152.16, 154.22, 156.45, 160.45, 166.45. HRMS (EI): m/z[M+H] calcd for C₂₄H₂₂N₄O₃: 415.1765. Found: 415.1770.

Synthesis 1463-tert-butyl-N-(3-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)benzamide(AA-029)

Method G1 was used with 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-2(1H)-oneand 3-tertbutylbenzoyl chloride to afford the title compound as a whitesolid (14 mg, 22%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.33 (s, 9H, tert-Bu), 6.94 (d, 1H,H_(Py), J=5.3 Hz), 7.00 (dd, 1H, H_(arom), J=8.1 Hz, J=2.4 Hz), 7.46 (t,1H, H_(arom), J=7.7 Hz), 7.49 (t, 1H, H_(arom), J=8.1 Hz), 7.63 (d, 1H,H_(arom), J=1.7 Hz), 7.71 (d, 1H, H_(arom), J=8.2 Hz), 7.75-7.77 (m, 2H,H_(arom)), 7.91 (t, 1H, H_(arom), J=7.9 Hz), 8.39 (d, 1H, H_(Py), J=5.3Hz), 8.43 (s, 1H, H_(arom)), 10.37 (s, 1H, NH_(amide)), 12.60 (s, 1H,NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 30.99 (tert-Bu),34.55 (tert-Bu), 110.60, 111.94, 115.27, 117.23, 117.99, 124.29, 124.79,128.11, 128.67, 130.27, 134.40, 134.96, 141.00, 150.94, 153.95, 154.77,155.88, 166.10. HRMS (EI): m/z [M+H] calcd for C₂₄H₂₂N₄O₃: 415.1765.Found: 415.1775.

Synthesis 147N-(3-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)-3-(trifluoromethoxy)benzamide (AA-030)

Method G1 was used with 8-(3-aminophenoxy)pyrido[2,3-b]pyrazin-2(1H)-oneand 3-trifluoromethoxybenzoyl chloride to afford the title compound as awhite solid (35 mg, 20%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.94 (d, 1H, H_(Py), J=5.3 Hz), 7.03(dd, 1H, H_(arom), J=8.1 Hz, J=2.3 Hz), 7.50 (t, 1H, H_(arom), J=8.2Hz), 7.61-7.62 (m, 1H, H_(arom)), 7.68-7.71 (m, 2H, H_(arom)), 7.75 (t,1H, H_(arom), J=2.0 Hz), 7.90 (s, 1H, H_(arom)), 8.00 (d, 1H, H_(arom),J=7.7 Hz), 8.39 (d, 1H, H_(Py) J=5.3 Hz), 8.43 (s, 1H, H_(arom),), 10.54(s, 1H, NHamide), 12.61 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ(ppm), J (Hz): 110.64, 111.98, 115.65, 117.27, 119.00, 120.17, 121.04,123.09, 124.18, 126.78, 130.40, 130.65, 136.75, 140.58, 145.36, 148.26,154.01, 154.61, 164.05. HRMS (EI): m/z [M+H] calcd for C₂₁H₁₃F₃N₄O₄:443.0962. Found: 443.0966.

Synthesis 148N-(3-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)-3-(trifluoromethoxy)benzamide)(AA-004)

Method G1 was used with8-(3-aminophenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one and3-trifluoromethoxybenzoyl chloride to afford the title compound as aslightly yellow solid (74 mg, 87%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 2.44 (s, 3H, Me), 6.62 (d, 1H,H_(Py), J=5.6 Hz), 7.01 (dd, 1H, H_(arom), J=8.1 Hz, J=2.3 Hz), 7.50 (t,1H, H_(arom), J=8.2 Hz), 7.61 (d, 1H, H_(arom), J=8.4 Hz), 7.67-7.71 (m,2H, H_(arom)), 7.73 (t, 1H, H_(arom), J=2.1 Hz), 7.90 (s, 1H, H_(arom)),8.01 (d, 1H, H_(arom), J=7.9 Hz), 8.31 (d, 1H, H_(Py) J=5.6 Hz), 10.55(s, 1H, NH_(amide)), 12.77 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ(ppm), J (HZ): 20.52 (Me), 106.55, 112.11, 115.72, 117.24, 117.91,120.03 (OCF₃), 120.23, 124.16, 126.85, 130.44, 130.62, 136.75, 140.66,145.76, 148.26, 150.58, 154.25, 156.32, 159.24, 159.65, 164.08. HRMS(EI): m/z [M+H] calcd for C₂₂H₁₅F₃N₄O₄: 457.1124. Found: 457.1118.

Synthesis 1493-tert-butyl-N-(3-(2-methyl-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)benzamide(AA-005)

Method G1 was used with8-(3-aminophenoxy)-2-methylpyrido[2,3-b]pyrazin-3(4H)-one and3-tertbutylbenzoyl chloride to afford the title compound as a slightlyyellow solid (77 mg, 97%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.33 (s, 9H, tert-Bu), 2.44 (s, 3H,Me), 6.62 (d, 1H, H_(Py), J=5.6 Hz), 6.98 (dd, 1H, H_(arom), J=8.1 Hz,J=2.4 Hz), 7.44-7.50 (m, 2H, H_(arom)), 7.63 (d, 1H, H_(arom), J=7.9Hz), 7.70 (d, 1H, H_(arom), J=8.2 Hz), 7.74-7.77 (m, 2H, H_(arom)), 7.91(s, 1H, H_(arom)), 8.30 (d, 1H, H_(Py) J=5.6 Hz), 10.37 (s, 1H,NH_(amide)), 12.77 (s, 1H, NH_(lactame)). ¹³C-NMR (DMSO-d6), δ (ppm), J(Hz): 20.52 (CH₃), 31.02 (tert-Bu), 34.57 (tert-Bu), 106.50, 112.06,115.36, 117.20, 117.88, 124.36, 124.85, 128.11, 128.67, 130.34, 134.43,141.07, 145.75, 150.58, 150.94, 154.19, 156.32, 159.20, 159.72, 166.14.HRMS (EI): m/z [M+H] calcd for C₂₅H₂₄N₄O₃: 429.1921. Found: 429.1921.

(VIII) Synthesis of Reverse Amides 1. Reverse Amides from CommonIntermediates Synthesis 150N-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)benzamide(AA-001)

Method H1: A solution of AlMe₃ (solution in toluene 2M, 0.85 mL, 1.68mmol,) was added dropwise to a cooled (0 C) solution of3-tert-butyl-1-phenyl-1H-pyrazol-5-amine (362 mg, 1.68 mmol) in THF (5.0mL). When the addition was complete, the mixture was allowed to warm toroom temperature and stirring was continued for 30 minutes. Then methyl3-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)benzoate (100 mg, 0.336mmol) was added and the mixture was heated under reflux for 19 h. Themixture was cooled to room temperature and carefully quenched with 5% aqHCl (3.0 mL). After evaporation of solvent, the residue was retaken inCH₂Cl₂, washed with saturated solution of NaHCO₃ and dried over MgSO₄and evaporated under vacuum. The obtained residue was chromatographied(eluent: CH₂Cl₂/EtOAc: 2/1 towards 1/3) and the title compound wasobtained as a slightly yellow solid (29 mg, 18%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.31 (s, 9H, tert-Bu), 6.39 (s, 1H,H_(Pyz)), 6.71 (d, 1H, H_(Py), J=5.6 Hz), 7.29-7.32 (m, 1H, H_(arom)),7.41-7.44 (m, 2H, H_(arom)), 7.47 (dd, 1H, H_(arom), J=8.1 Hz, J=2.4Hz), 7.50-7.52 (m, 2H, H_(arom)), 7.61-7.64 (m, 2H, H_(arom)), 7.78 (d,1H, H_(arom), J=7.7 Hz), 8.17 (s, 1H, H-_(arom)), 8.42 (d, 1H, H_(Py),J=5.6 Hz), 10.35 (s, 1H, arom, NH_(amide)), 12.94 (s, 1H, NH_(lactame)).¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 30.03 (tert-Bu), 32.00 (tert-Bu),100.70, 107.39, 118.62, 118.67, 122.92, 122.98, 123.45, 124.39, 126.75,128.87, 130.68, 135.32, 138.88, 145.58, 151.30, 152.17, 154.50, 156.35,159.73, 160.71, 164.71. HRMS (EI): m/z [M+H] calcd for C₂₇H₂₄N₆O₃:481.1983. Found: 481.1983.

(IX) Synthesis of Ureas from Isocyanates and Nitro-Amino-PyridineIntermediates Synthesis 1511-(4-(2-amino-3-nitropyridin-4-yloxy)-2-(methyltio)phenyl)-3-(4-chloro-3-(trifluoromethyl)phenyl)urea

Using Method F2 with4-(4-amino-3-(methylthio)phenoxy)-3-nitropyridin-2-amine (150 mg, 0.5mmol) and 4-chloro-3-trifluoromethylisocyanate, the title compound (247mg, 93%) was obtained as a orange powder.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 2.47 (5, 3H, CH₃), 6.02 (d, 1H,H_(Py) J=5.7 Hz), 7.04 (d, 1H, H_(arom), J=8.8 Hz), 7.16 (5, 2H,NH_(2,Py)), 7.21 (m, 1H, H_(arom), J=8.8 Hz), 7.62 (m, 2H, H_(arom)),7.85 (m, 1H, H_(arom)), 8.01 (d, 1H, H_(arom), J=8.8 Hz), 8.11 (d, 1H,H_(Py) J=5.7 Hz), 8.20 (s, 1H, NH_(urea1)), 9.75 (s, 1H, NH_(urea3)).¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 15.6, 100.4, 116.5, 118.0, 119.8,121.6, 122.7, 123.8, 124.0, 126.5, 126.8, 131.7, 132.0, 133.9, 139.2,149.3, 152.4, 153.1, 153.7, 158.9. LC-MS (m/z): 514 (M+H, 100), rt=8.37min.

Synthesis 1521-(4-(2-amino-3-nitropyridin-4-yloxy)-2-(methylthio)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea

Using Method F2 with4-(4-amino-3-(methylthio)phenoxy)-3-nitropyridin-2-amine (1.04 g, 3.57mmol) and 2-fluoro-5-trifluoromethylphenyl isocyanate, the titlecompound (664 mg, 37%) was obtained as a yellow powder.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 2.48 (s, 3H, CH₃), 6.02 (d, 1H,H_(Py) J=5.7 Hz), 7.02 (dd, 1H, H_(arom), J=8.7 Hz, J=2.7 Hz), 7.39 (m,1H, H_(arom)), 7.50 (m, 1H, H_(arom)), 7.83 (d, 1H, H_(arom), J=8.8 Hz),8.01 (d, 1H, H_(Py) J=5.7 Hz), 8.62 (dd, 1H, H_(arom), J=7.1 Hz, J=1.6Hz), 8.66 (s, 1H, H_(arom)), 9.69 (s, 1H, NH_(urea1)), 10.50 (s, 1H,NH_(urea3)). LC-MS (m/z): 498 (M+H, 100), rt=5.54 min.

Synthesis 1531-(4-(2-amino-3-nitropyridin-4-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoro-methyl)phenyl)urea

Using Method F2 with 4-(4-amino-3-fluorophenoxy)-3-nitropyridin-2-amineand 2-fluoro-5-trifluoromethylphenyl isocyanate, the title compound wasobtained (yield 85%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.03 (d, 1H, J=5.7 Hz), 7.04 (dd, 1H,J=8.6, 2.2 Hz), 7.22 (bs, 2H), 7.33 (dd, 1H, J=8.6, 2.9 Hz), 8.60 (m,1H), 9.22 (s, 1H), 9.37 (s, 1H). LC-MS (m/z): 470 (M+H, 100).

Synthesis 1541-(4-(2-amino-3-nitropyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)urea

Using Method F3 with -tert-butyl-5-isocyanato-1-phenyl-1H-pyrazole (15mL, 4.05 mmol) and 4-(4-amino-3-fluorophenoxy)-3-nitropyridin-2-amine(893 mg, 3.38 mmol) the title compound was obtained in quantitive yield(1.71 g) as a yellow solid after column chromatography with 5% to 50%EtOAc in CH₂Cl₂.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 6.03 (d, 1H,J=5.7, H_(Py)), 6.40 (s, 1H, H_(Pyz)), 7.01 (m, 1H, H_(arom)), 7.18 (brs, 2H, NH₂), 7.26 (m, 1H, H_(arom)), 7.43 (m, 1H, H_(arom)), 7.54 (m,4H, H_(arom)), 8.01 (d, 1H, J=5.7, H_(Py)), 8.16 (m, 1H, H_(arom)), 8.84(s, 1H, NH), 8.98 (br s, 1H, NH); ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz):30.1, 32.0, 95.1, 100.6, 108.6 (d, J_(FC)=22.6), 116.6, 124.4, 125.2 (d,J_(FC)=10.8), 127.3, 129.3, 136.9, 138.4, 147.7 (d, J_(FC)=10.4), 151.1,152.1 (d, J_(FC)=246), 153.2, 153.9, 158.8, 160.8, 170.3;

¹⁹F-NMR (DMSO-d6), δ (ppm): −124.7; LC-MS (m/z): 506.1 (M+H, 100),rt=2.73 min.

(X) Reduction of Nitro Group of Coupled Intermediates (According toScheme 9) Synthesis 1551-(4-(2,3-Diaminopyridin-4-yloxy)-2-(methylthio)phenyl)-3-(4-chloro-3-(trifluoromethyl)phenyl)urea

Method C4. A suspension of iron powder (4 equivalents, 78 mg, 1.4 mmol)and ammonium chloride (5.8 equivalents, 109 mg, 2 mmol) in ethanol (400μL) and water (438 μL) was heated to reflux. The1-(4-(2-amino-3-nitropyridin-4-yloxy)-2-(methyltio)phenyl)-3-(4-chloro-3-(trifluoromethyl)phenyl)ureacompound (180 mg, 0.35 mmol) was added in portions and the mixturestirred at reflux for 24 hours. After cooling at RT, the slurry mixturewas filtered and washed with ethanol. After removed the solvent, thecrude powder is dissolved into EtOAc, filtered to removed theprecipitate, and evaporated to provide the title compound (100 mg, 59%)as a sticky dark oil.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 2.41 (s, 3H, CH₃), 5.61 (s, 2H,NH_(2,Py)), 6.06 (d, 1H, H_(Py) J=5.6 Hz), 6.79 (d, 1H, H_(arom), J=8.7Hz), 7.01 (s, 1H, H_(arom)), 7.26 (d, 1H, H_(Py), J=5.6 Hz), 7.58-7.69(m, 4H, H_(arom)), 8.12 (s, 2H, NH_(2, Py)), 8.27 (s, 1H, NH_(urea1)),10.02 (s, 1H, NH_(urea3)). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 15.6,103.8, 115.6, 116.3, 117.6, 119.9, 122.5, 122.6, 124.9, 131.7, 131.9,132.4, 134.7, 139.5, 139.6, 144.1, 147.0, 149.9, 152.3, 152.8. LC-MS(m/z): 484 (M+H, 100), rt=5.81 min.

Synthesis 1561-(4-(2,3-diaminopyridin-4-yloxy)-2-(methylthio)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea

Using Method C4 with1-(4-(2-amino-3-nitropyridin-4-yloxy)-2-(methylthio)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(664 mg, 1.3 mmol), the title compound (120 mg, 19%) was obtained as adark powder after purification by chromatography on silica gel (EtOAc,then EtOAc-MeOH: 95-5)(R_(f) 0.33, EtOAc-MeOH, 95:5).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 2.43 (s, 3H, CH₃), 4.45 (s, 2H,NH_(2,Py)), 5.57 (s, 2H, NH_(2,Py)), 6.07 (d, 1H, H_(Py) J=5.6 Hz), 6.79(dd, 1H, H_(arom), J=8.7 Hz, J=2.7 Hz), 7.01 (d, 1H, H_(arom), J=2.7Hz), 7.27 (d, 1H, H_(Py) J=5.6 Hz), 7.37 (m, 1H, H_(arom)), 7.49 (m, 1H,H_(arom)), 7.67 (d, 1H, H_(arom), J=8.8 Hz), 8.57 (s, 1H, NH_(urea1)),8.62 (dd, 1H, H_(arom), J=7.3 Hz, J=2.0 Hz), 9.43 (s, 1H, NH_(urea3)).¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 15.4, 103.8, 115.4, 115.9, 116.0,116.6, 117.3, 119.0, 119.8, 124.9, 128.6, 128.7, 131.3, 131.9, 135.5,146.8, 150.2, 152.4, 152.5, 154.3. LC-MS (m/z): 468 (M+H, 100), rt=3.48min.

Synthesis 1571-(4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea

Method C2 was used with1-(4-(2-amino-3-nitropyridin-4-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(500 mg, 1.08 mmol) to yield the title compound (450 mg, 95%) as ayellow solid.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 5.38 (bs, 2H), 6.05 (d, 1H, J=5.9Hz), 6.75-6.86 (m, 2H), 7.21-7.33 (m, 4H) 8.07 (dd, 1H, J=18.0, 9.7 Hz),8.94 (bs, 1H), 9.15 (bs, 1H). LC-MS (m/z): 440 (M+H, 100).

Synthesis 1581-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenyl)urea

Method C2 was used with1-(4-(2-amino-3-nitropyridin-4-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)urea(810 mg, 1.60 mmol) to give the title compound as a light pink solid(750 mg, 99% yield).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.28 (s, 9H, tert-Bu), 4.45 (br s,2H, NH₂), 5.58 (br s, 2H, NH₂), 6.06 (d, 1H, J=5.6, H_(Py)), 6.38 (s,1H, H_(Pyz)), 6.78 (m, 1H, H_(arom)), 6.92 (m, 1H, H_(arom)), 7.26 (d,1H, J=5.6, H_(Py)), 7.41 (m, 1H, H_(arom)), 7.52 (m, 4H, H_(arom)), 7.98(m, 1H, H_(arom)), 8.74 (s, 1H, NH), 8.82 (br s, 1H, NH); LC-MS (2.19min): m/z 476.2 (M+H, 100).

(XI) Cyclisation of Coupled Intermediates Synthesis 1591-(4-chloro-3-(trifluoromethyl)phenyl)-3-(4-(2,3-dioxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenyl)urea(AA-051)

Method D3 was used with1-(4-(2,3-diaminopyridin-4-yloxy)-2-(methylthio)phenyl)-3-(4-chloro-3-(trifluoromethyl)phenyl)urea(65 mg, 0.1 mmol) to provide the title compound (9 mg, 12%) as a palewhite powder.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 2.46 (s, 3H, CH₃); 6.55 (d, 1H,H_(Py) J=5.5 Hz), 7.03 (dd, 1H, H_(arom), J=8.7 Hz, J=2.5 Hz), 7.21 (d,1H, H_(arom), J=2.6 Hz), 7.62 (m, 2H, H_(arom)), 7.83 (d, 1H, H_(arom),J=8.7 Hz), 7.95 (d, 1H, H_(Py) J=5.0 Hz), 8.11 (m, 1H, H_(arom)), 8.22(s, 1H, NH or CH), 9.81 (s, 1H, NH or CH), 11.89 (s, 1H, NH or CH),12.38 (s, 1H, NH). ¹³C-NMR (DMSO-d6), δ (ppm), J (Hz): 15.7, 106.2,112.2, 116.5, 117.7, 119.6, 122.2, 122.7, 122.8, 124.1, 129.6, 131.6,131.9, 133.4, 139.2, 140.4, 143.1, 150.1, 150.4, 152.4, 15.5, 155.8.LC-MS (m/z): 538 (M+H, 100), rt=4.98 min.

Synthesis 1601-(4-(2,3-dioxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-8-yloxy)-2-(methylthio)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-052)

Method D3 was used with1-(4-(2,3-diaminopyridin-4-yloxy)-2-(methylthio)phenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(87 mg, 0.18 mmol) to afford the title compound (34 mg, 35%) wasobtained as a powder.

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 2.47 (s, 3H, CH₃), 6.56 (d, 1H,H_(Py) J=5.6 Hz), 7.02 (dd, 1H, H_(arom), J=8.7 Hz, J=1.7 Hz), 7.19 (d,1H, H_(arom), J=1.7 Hz), 7.39 (m, 1H, H_(arom)), 7.50 (m, 1H, H_(arom)),7.82 (d, 1H, H_(arom), J=8.7 Hz), 7.95 (dd, 1H, H_(arom), J=5.6 Hz,J=0.9 Hz), 8.63 (d, 1H, H_(Py) J=6.7 Hz), 8.67 (s, 1H, NH), 9.52 (s, 1H,NH), 11.90 (s, 1H, NH), 12.39 (s, 1H, NH). ¹³C-NMR (DMSO-d6), δ (ppm), J(Hz): 15.5, 106.2, 112.2, 115.9, 116.1, 116.7, 117.4, 119.2, 122.7,124.6, 125.1, 128.6, 128.7, 131.9, 133.0, 140.4, 143.1, 150.3, 150.4,152.4, 154.6, 155.8. LC-MS (m/z): 522 (M+H, 100), rt=4.82 min.

Synthesis 1611-(4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)-3-(4-chloro-3-(trifluoromethyl)phenyl)urea(AA-021)

Method D4 was used with1-(4-chloro-3-(trifluoromethyl)phenyl)-3-(4-(2,3-diaminopyridin-4-yloxy)phenyl)urea(50 mg, 0.12 mmol) to afford the title compound (10 mg, 17% yield) as awhite solid.

¹H-NMR (CD₃OD), δ (ppm), J (Hz): 8.03 (m, 2H), 8.68-8.73 (m, 4H), 8.94(dd, 1H, J=8.8, 2.6 Hz), 9.58 (m, 2H), 10.86 (bs, 1H), 11.84 (bs, 1H).LC-MS (m/z): 491.0 (M+H, 100).

Synthesis 1621-(2-fluoro-5-(trifluoromethyl)phenyl)-3-(4-(2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)urea(AA-043)

Method D1 was used with1-(4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)ureato afford the title compound (yield 32%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.70 (d, 1H, J=5.4 Hz), 7.23 (d, 2H,J=9.6 Hz), 7.39 (m, 1H), 7. 50 (m, 1H), 7.60 (d, 2H, J=9.6 Hz), 8.34 (d,1H, J=5.4), 8.41 (s, 1H), 8.61 (dd, 1H, J=7.4, 1.6 Hz), 12.52 (bs, 1H).LC-MS (m/z): 492 (M+H, 100).

Synthesis 1631-(4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-044)

Method D4 was used with1-(4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)ureato yield 40 mg (25%) of the title compound as a white solid.

¹H-NMR (DMSO), δ (ppm), J (Hz): 6.57 (d, 1H, 2H, J=5.6 Hz), 7.01 (dd,1H, J=11.7, 2.8 Hz), 7.48-7.52 (m, 2H), 8.10 (d, 1H, J=5.6 Hz), 8.15 (d,1H, 8.4 Hz), 8.60 (dd, 1H, J=8.4, 2.8 Hz), 9.34 (bs, 1H), 9.5 (bs, 1H).LC-MS (m/z): 493 (M+H, 100).

Synthesis 1641-(4-(3-amino-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)urea(AA-022)

Method D4 was used with1-(4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenyl)-3-(2-fluoro-5-(trifluoromethyl)phenyl)ureato yield the title compound (yield 38%).

¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 6.56 (d, 1H, J=5.4 Hz), 7.02 (dd, 1H,J=9.2, 2.7 Hz), 7.24 (dd, 1H, J=11.3, 2.6 Hz), 7.47-7.52 (m, 2H), 8.10(d, 1H, 5.4 Hz), 8.15 (m, 1H), 8.61 (dd, 1H, J=7.3, 2.5 Hz), 9.35 (bs,1H), 9.50 (bs, 1H). LC-MS (m/z): 493 (M+H, 100).

Synthesis 1651-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)ureaand1-(3-tea-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-oxo-1,2-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-019 and AA-089)

Method D1 was used with1-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenyl)urea(730 mg, 1.54 mmol) to give1-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)ureaas the first fraction (412 mg, 52%) and1-(3-tert-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-oxo-1,2-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)ureaas the second (300 mg, 38%).

1-(3-tea-butyl-1-phenyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-oxo-1,2-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea:¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.30 (s, 9H, tert-Bu), 6.41 (s, 1H,H_(Pyrazole)), 6.92 (d, 1H, J=5.4, H_(Py)), 7.08 (m, 1H, H_(arom)), 7.31(m, 1H, H_(arom)), 7.44 (m, 1H, H_(arom)), 7.55 (m, 4H, H_(arom)), 8.18(m, 1H, H_(arom)), 8.37 (d, 1H, J=5.4, H_(Py)), 8.43 (s, 1H, H_(arom)),8.85 (s, 1H, NH), 9.01 (br s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J(Hz): 30.2, 32.0, 95.1, 99.5, 108.6 (d, J_(FC)=22.5), 116.5, 121.7,124.4, 124.9 (d, J_(FC)=10.8), 127.4, 129.3, 135.1, 136.9, 138.4, 139.5,145.3 (br), 148.4 (d, J_(FC)=10.4), 149.7, 151.4, 152.2 (d, J_(FC)=248),160.8; ¹⁹F-NMR (DMSO-d₆), δ (ppm): −124.7; LC-MS (m/z): 514.1 (M+H,100), rt=2.54 min; HRMS (3.10 min): m/z calcd. for C₂₇H₂₅FN₇O₃ (M+H,100)⁺: 514.19974. Found: 514.19856.

Synthesis 1661-(4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea(AA-057) and1-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-oxo-1,2-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-085)

Method D4 was used with1-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenyl)urea(250 mg, 0.51 mmol) to afford after chromatography1-(4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea(AA-057)(25 mg, 9% yield) and1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-oxo-1,2-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-085) (15 mg, 6% yield).

1-(4-(2-amino-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea(AA-057): ¹H-NMR (CD3OD), δ (ppm), J (Hz): 1.36 (s, 9H), 6.46 (s, 1H),6.65 (d, 1H, J=5.7 Hz), 6.97 (d, 1H, J=9.0 Hz), 7.04 (dd, 1H, J=9.0, 2.6Hz), 7.41 (AB system, 4H) 8.05 (d, 1H, J=5.7 Hz), 8.11 (t, 1H, J=9.0Hz), 8.79 (bs, 1H), 9.00 (bs, 1H), 11.24 (bs, 1H), 12.26 (bs, 1H). LC-MS(m/z): 544 (M+H, 100).

1-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-oxo-1,2-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-085): ¹H-NMR (DMSO-d6), δ (ppm), J (Hz): 1.31-1.28 (m, 9H), 3.33 (s,3H), 6.43 (s, 1H), 6.66 (d, 1H, J=5.6 Hz), 7.05 (dd, 1H, J=8.1, 2.0 Hz),7.31 (dd, 1H, J=11.8, 2.0 Hz), 7.45 (d, 1H, J=8.3 Hz), 7.85 (dd, 1H,J=8.0, 3.3 Hz), 8.15 (t, 1H, J=9.2 Hz), 8.18 (s, 1H), 8.38 (d, 1H, J=6.0Hz), 8.62 (d, 1H, J=3.3 Hz), 8.90 (s, 1H), 8.98 (s, 1H), 12.93 (bs, 1H).LC-MS: 544 (M+H, 100). HRMS: m/z calcd. for C27H25FN8O (M+H, 100):543.2263. Found: 543.2262.

Synthesis 1671-(4-(3-(bromomethyl)-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea(AA-058) and1-(4-(2-(bromomethyl)-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea(AA-059)

Method D8: To a solution of1-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)-3-(4-(2,3-diaminopyridin-4-yloxy)-2-fluorophenyl)urea(300 g, 0.61 mol) in dry ethanol (5 ml) ethyl 3-bromo-2-oxopropanoate(390 mg, 2 mmol) was added in one go. The resulting suspension wasrefluxed for 4 days. The solvent was then evaporated and chromatographedon a Biotage apparatus to afford 24 mg (6% yield) of1-(4-(3-(bromomethyl)-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureaand 13 mg (4% yield) of1-(4-(2-(bromomethyl)-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea.

1-(4-(3-(bromomethyl)-2-oxo-1,2-dihydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea(AA-058): ¹H-NMR (CD3OD), δ (ppm), J (Hz): 1.34 (s, 9H), 2.42 (s, 3H),4.63 (s, 2H), 6.46 (s, 1H), 6.66 (d, 1H, J=5.6 Hz), 7.05 (1H, dd, J=9.0,2.5 Hz), 7.13 (1H, dd, J=9.0, 2.5 Hz), 7.37-7.35 (AB, 4H), 8.16 (t, 1H,J=9.0), 8.34 (d, 1H, J=5.6 Hz); LC-MS (m/z): 622-620 (M+H, 100).

1-(4-(2-(bromomethyl)-3-oxo-1,2,3,4-tetrahydropyrido[2,3-b]pyrazin-8-yloxy)-2-fluorophenyl)-3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)urea(AA-059): ¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.27 (s, 9H), 2.08 (s, 3H),2.39 (s, 2H), 6.92-6.90 (m, 2H), 7.12 (m, 1H), 7.13 (m, 1H), 7.38-7.35(4H, AB), 7.92 (1H, d, J=5.7 Hz), 8.09 (1H, dd, J=4.7, 5.0 Hz), 8.74(1H, s), 8.92 (1H, bs); LC-MS (m/z): 622-620 (M+H, 100).

(XII) Synthesis of Ureas from Activated Carbamates and AminoIntermediates Synthesis 1681-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenyl)-3-(5-(trifluoromethyl)pyridin-3-yl)urea(AA-069)

Method F5. w 8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one(26 mg, 96 μmol) and prop-1-en-2-yl5-(trifluoromethyl)pyridin-3-ylcarbamate (45.8 mg, 186 μmol) wereweighed into a 10 mL RBF, put under Ar and dry THF (3 mL) was added. Tothis mixture, N-methylpyrrolidine (1 drop) was added and the mixture washeated to reflux for 48 h. The volatiles were evaporated and theresulting mixture was re-dissolved in MeOH (3 mL) and evaporated ontosilica gel, which was loaded onto a silica gel column and purified witha 0-20% MeOH in EtOAc gradient. Yield: 5 mg (11%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.66 (d, J=5.6, 1H, H_(Py)), 7.08 (m,1H, H_(arom)), 7.35 (m, 1H, H_(arom)), 8.13 (m, 1H, H_(arom)), 8.18 (s,1H, H_(arom)), 8.38 (d, J=5.6, 1H, H_(Py)), 8.46 (s, 1H, H_(arom)), 8.59(s, 1H, H_(arom)), 8.77 (s, 1H, H_(arom)), 8.96 (s, 1H, NH), 9.67 (s,1H, NH), 12.95 (s, 1H, NH); ¹⁹F-NMR (DMSO-d₆), δ (ppm): −60.6, −123.7;LC-MS (m/z): LC-MS: 461.1 (M+H, 100), rt=2.44 min; HRMS (7.17 min): m/zcalcd. for C₂₀H₁₃F₄N₆O₃ [M+H⁺]: 461.09798. Found: 461.09771.

Synthesis 169 prop-1-en-2-yl 5-(trifluoromethyl)pyridin-3-ylcarbamate

5-(trifluoromethyl)pyridin-3-amine (883 mg, 5.45 mmol) was suspended indry THF (20 mL) and N-methylpyrrolidine (680 μL, 6.54 mmol) was added togive a brown suspension. The mixture was cooled to 0° C. and isopropenylchloroformate (715 μL, 6.54 mmol) was added dropwise over 15 min. Thesuspension was allowed to reach RT and was stirred for 4 h. EtOAc (60mL) and H₂O (10 mL) were added and the organic layer was isolated,washed with 50% brine (10 mL), dried (MgSO₄), filtered and evaporated toleave a brown oil, which solidified upon standing (1.05 g). The solidwas taken up in CH₂Cl₂ (4 mL) and purified by column chromatography onsilica gel, eluting with EtOAc in CH₂Cl₂ (6%→40%), to give a whitesolid. Yield: 600 mg (45%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.96 (s, 3H, CH₃), 4.78 (m, 1H, CH),4.80 (m, 1H, CH), 8.27 (br, 1H, H_(arom)), 8.62 (br, 1H, H_(arom)), 8.86(d, J=5.5, 1H, H_(Py)), 10.54 (s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J(Hz): 19.3, 102.1, 121.2, 123.5 (q, J_(FC)=272), 125.1 (q, J_(FC)=31),135.8, 139.8 (q, J_(FC)=3.8), 143.7, 151.3, 152.2; ¹⁹F-NMR (DMSO-d₆), δ(ppm): −61.2; LC-MS (m/z): m/z 247.0 (M+H, 100), rt=4.49 min.

Synthesis 170 Phenyl2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenylcarbamate

Dry pyridine (125 μL, 1.55 mmol) was added to a suspension of8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one (307 mg, 1.13mmol) in dry THF (20 mL) under Ar and the mixture was cooled to 0° C.Phenyl chloroformate (170 μL, 1.35 mmol) was added dropwise over 5 minand the light brown mixture was stirred at 0° C. for an additional 5 minwhereafter the mixture was allowed to warm up to RT and was stirred for150 min. The brownish mixture was concentrated to dryness and theresulting residue was diluted with EtOAc (60 mL) and H₂O (30 mL). Theorganic layer was isolated and filtered (80 mg of impure product) andthe filtrate was washed with aqueous saturated NaHCO₃ and brine. Theorganic layer was evaporated to dryness, re-dissolved in CH₂Cl₂ andchromatographed on a Biotage 25+M column, eluting with 20%100% EtOAc inCH₂Cl₂ to give the title compound as a white solid. Yield: 280 mg (81%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.71 (d, J=5.6, 1H, H_(Py)), 7.09 (m,1H, H_(arom)), 7.23 (d, J=7.9, 2H, H_(arom)), 7.26 (t, J=7.9, 1H,H_(arom)), 7.33 (m, 1H, H_(arom)), 7.43 (d, J=7.9, 2H, H_(arom)), 7.75(m, 1H, H_(arom)), 8.18 (s, 1H, H_(arom)), 8.39 (d, J=5.6, 1H, H_(Py)),10.06 (s, 1H, NHBoc), 12.97 (s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J(Hz): 107.1, 108.6 (d, J_(FC)=22.9), 116.1 (d, J_(FC)=3.3), 118.6,121.8, 122.9 (d, J_(FC)=12.0), 125.5, 125.6 (br), 129.4, 145.6, 150.6,151.3, 151.4 (br), 152.3, 152.4, 154.8 (d, J_(FC)=245), 156.4, 160.0;¹⁹F-NMR (DMSO-d₆), δ (ppm): −119.2; LC-MS (m/z): 393.1 (M+H, 100),rt=2.44 min.

Synthesis 171 1-(2-nitro-4-(trifluoromethyl)phenyl)-1H-imidazole

A mixture of imidazole (0.997 g, 14.65 mmol) and tert-BuOK (1.722 g,15.35 mmol) was put under Ar in a 100 mL and dissolved in dry DMSO (15mL) to give a colorless solution. After 5 min,1-fluoro-2-nitro-4-(trifluoromethyl)benzene (2.04 mL, 14.58 mmol) wasadded within 30 s, immediately leading to a darkening of the rm toblack. A temperature rise was also noted. The black solution was stirredat RT for 20 min. Ice water (60 mL) and EtOAc (50 mL) were added, theorganic layer was isolated, and the aqueous phase was extracted twicewith 20 mL EtOAc. The organic layer was washed with H₂O (2×30 mL),brine, dried, filtered and evaporated to give the title compound as anorange oil. Yield: 3.66 g (97%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 7.14 (s, 1H, H_(arom)), 7.49 (s, 1H,H_(arom)), 7.97 (d, J=8.4, 1H, H_(arom)), 7.99 (s, 1H, H_(arom)), 8.28(d, J=8.4, 1H, H_(arom)), 8.59 (s, 1H, H_(arom))_(;) ¹³C-NMR (DMSO-d₆),δ (ppm), J (Hz): 120.4, 122.7 (d, J_(FC)=274), 122.9, 129.5 (d,J_(FC)=34), 129.9, 130.0, 130.9, 133.4, 137.4, 144.5; ¹⁹F-NMR (DMSO-d₆),δ (ppm): −60.8; LC-MS (m/z): 258.1 (M+H, 100), rt=1.37 min.

Synthesis 172 2-(1H-pyrazol-1-yl)-5-(trifluoromethyl)aniline

Method C3 was used 1-(2-nitro-4-(trifluoromethyl)phenyl)-1H-pyrazole(1.80 g, 7.00 mmol) in EtOH (40 mL) to give 760 mg (48%) of the titlecompound as white crystals after crystallization from hexane.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.12 (s, 2H, NH₂), 6.56 (vt, J=2.1,1H, H_(arom)), 6.95 (dd, J=8.3, ⁴J_(FH)=1.7, 1H, H_(arom)), 7.24 (d,⁴J_(FH)=1.7, 1H, H_(arom)), 7.48 (d, J=8.3, 1H, H_(arom)), 7.82 (d,J=1.8, 1H, H_(arom)), 8.23 (d, J=2.5, 1H, H_(arom)); ¹³C-NMR (DMSO-d₆),δ (ppm), J (Hz): 106.9, 112.1, 112.9, 124.1 (d, J_(FC)=273), 124.2,127.4, 128.3 (d, J_(FC)=31.7), 130.6, 140.5, 142.1; ¹⁹F-NMR (DMSO-d₆), δ(ppm): −60.8.

Synthesis 173 3-(2-nitro-4-(trifluoromethyl)phenoxy)pyridine

A brown solution of 1-fluoro-2-nitro-4-(trifluoromethyl)benzene (2.01 g,9.61 mmol) and 3-hydroxypyridine (0.923 g, 9.71 mmol) in dry DMF (15 ml)under Ar was treated with cesium carbonate (3.28 g, 10.07 mmol) at onceand the brown mixture was stirred at RT for 2 h. H₂O (50 mL) and EtOAc(50 mL) were added and the organic layer was isolated. The water layerwas extracted with EtOAc (2×30 mL). The combined organic layer waswashed with H₂O (3×40 mL), brine (40 mL), dried (MgSO₄), filtered andconcentrated to dryness to give a light yellow solid. Yield: 2.62 g(96%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 7.33 (d, J=8.7, 1H, H_(arom)), 7.53(m, 1H, H_(arom)), 7.71 (m, 1H, H_(arom)), 8.04 (dd, J=8.9, ⁴J_(FH)=2.3,1H, H_(arom)), 8.49 (d, ⁴J_(FH)=2.2, 1H, H_(arom)), 8.52 (m, 1H,H_(arom)), 8.55 (d, J=2.9, 1H, H_(arom)); ¹³C-NMR (DMSO-d₆), δ (ppm), J(Hz): 120.7, 122.9 (d, J_(FC)=274), 123.4, 124.3 (d, J_(FC)=33.9),125.0, 127.2, 131.7, 140.6, 141.6, 146.6, 151.3, 152.1; ¹⁹F-NMR(DMSO-d₆), δ (ppm): −60.4; LC-MS (m/z): 285.0 (M+H, 100), rt=2.40 min.

Synthesis 174 2-(pyridin-3-yloxy)-5-(trifluoromethyl)aniline

Method C3 was used with 3-(2-nitro-4-(trifluoromethyl)phenoxy)pyridine(594 mg, 2.090 mmol) to give the title compound as a white crystallinesolid. Yield: 501 mg (94%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 5.55 (s, 2H, NH₂), 6.83 (d, J=8.2,1H, H_(arom)), 6.94 (d, J=8.2, 1H, H_(arom)), 7.13 (s, 1H, H_(arom)),7.33 (m, 1H, H_(arom)), 7.39 (m, 1H, H_(arom)), 8.33 (m, 1H, H_(arom)),8.37 (m, 1H, H_(arom)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 111.7,112.5, 119.7, 124.3 (d, J_(FC)=274), 124.4, 124.5, 125.8 (d,J_(FC)=33.9), 140.2, 141.0, 144.0, 144.1, 153.0; ¹⁹F-NMR (DMSO-d₆), δ(ppm): −60.3; LC-MS m/z: 255.0 (M+H, 100), rt=2.26 min.

Synthesis 175 Phenyl2-(pyridin-3-yloxy)-5-(trifluoromethyl)phenylcarbamate

A yellow solution of 2-(pyridin-3-yloxy)-5-(trifluoromethyl)aniline (263mg, 1.035 mmol) and pyridine (108 μL, 1.341 mmol) in dry THF (8 mL) wastreated dropwise with phenyl chloroformate (156 μL, 1.242 mmol) during 5min at 0° C. The resulting suspension yellow suspension was stirred at0° C. for an additional 5 min and then allowed to warm up to roomtemperature and stirred for 3 h. The yellow suspension was filtered overcotton, washed with Et₂O and diluted with EtOAc. The yellow solution waswashed with sat. aqueous NaHCO₃ (30 mL) and H₂O (30 mL), dried andconcentrated to dryness to give a yellow oil. Purification by columngave a tan solid. Yield: 300 mg (77%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.90 (d, J=8.5, 1H, H_(arom)), 7.23(m, 2H, H_(arom)), 7.30 (m, 2H, H_(arom)), 7.42 (m, 4H, H_(arom)), 7.76(br s, 1H, H_(arom)), 8.55 (m, 1H, H_(arom)), 8.64 (br s, 1H, NH); LC-MS(m/z): 375.0 (M+H, 100), rt=2.62 min.

Synthesis 1761-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)-3-(2-(pyridin-3-yloxy)-5-(trifluoromethyl)phenyl)urea(AA-093)

Method F2 was used with8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H-one (30.6 mg, 0.112mmol) and a 60.6 mM solution of phenyl2-(pyridin-3-yloxy)-5-(trifluoromethyl)phenylcarbamate (1.6 ml, 0.097mmol). After 40 h, the mixture was evaporated onto silica gel, loadedonto a Biotage 12+M column, which was eluted with 40%-100% EtOAc in DCM.Yield: 4 mg (7%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.64 (d, J=5.6, 1H, H_(Py)), 7.02 (d,J=8.5, 1H, H_(arom)), 7.07 (m, 1H, H_(arom)), 7.32 (m, 2H, H_(arom)),7.53 (m, 1H, H_(arom)), 7.65 (m, 1H, H_(arom)), 8.15 (s, 1H, H_(arom)),8.26 (m, 1H, H_(arom)), 8.38 (d, J=5.6, 1H, H_(arom)), 8.50 (m, 1H,H_(arom)), 8.58 (d, J=2.8, 1H, H_(arom)), 8.74 (d, J=2.8, 1H, H_(arom)),9.36 (s, 1H, NH), 9.39 (s, 1H, NH), 12.88 (s, 1H, NH); LC-MS (m/z):553.1 (M+H, 100), rt=2.63 min; HRMS (3.22 min): m/z calcd. forC₂₆H₁₆F₄N₆O₄ (M+H, 100)⁺: 553.12419. Found: 553.12312.

Synthesis 1771-(2-(1H-pyrazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-094)

Method F4: A mixture of phenyl2-fluoro-4-(3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-8-yloxy)phenylcarbamate(36.3 mg, 0.093 mmol) and2-(1H-imidazol-1-yl)-5-(trifluoromethyl)aniline (21.1 mg, 0.093 mmol)was dissolved in dry DMSO (250 μL) the resulting orange solution wasstirred at 60° C. for 7 h. The solution was diluted with H₂O, extractedwith EtOAc and the organic layer was dried and evaporated to dryness.After a column (DCM/EtOAc) the resulting oil was triturated with EtOAcand the resulting white solid was collected. Yield: 11 mg (23%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 6.67 (m, 2H, H_(arom)), 7.07 (m, 1H,H_(arom)), 7.31 (m, 1H, H_(arom)), 7.53 (m, 1H, H_(arom)), 7.69 (m, 1H,H_(arom)), 7.93 (s, 1H, H_(arom)), 8.04 (m, 1H, H_(arom)), 8.19 (s, 1H,H_(arom)), 8.35 (m, 1H, H_(arom)), 8.39 (m, 1H, H_(arom)), 8.59 (s, 1H,H_(arom)), 9.40 (s, 1H, H_(arom)), 9.52 (s, 1H, H_(arom)), 12.93 (s, 1H,H_(arom)); ¹⁹F-NMR (DMSO-d₆), δ (ppm): −60.3, −122.1; LC-MS (m/z): 526.1(M+H, 100), rt=2.54 min; HRMS (3.10 min): m/z calcd. for C₂₄H₁₆F₄N₇O₃(M+H, 100)⁺: 526.12453. Found: 526.12498;

Synthesis 1781-(3-tert-butyl-1-(6-methylpyridin-3-yl)-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-084)

Method F4 was used with 65 mg (0.17 mmol) of phenyl2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenylcarbamateand 45 mg (0.2 mmol) of3-tert-butyl-1-(6-methylpyridin-3-yl)-1H-pyrazol-5-amine (Regan, J. etal., J. Med. Chem. 2002, 45, 2994-3008). Obtained 15 mg, 17% yield ofthe title compound.

¹H-NMR (CD3OD), δ (ppm), J (Hz): 1.31-1.28 (m, 9H), 3.33 (s, 3H), 6.43(s, 1H), 6.66 (d, 1H, J=5.6 Hz), 7.05 (dd, 1H, J=8.1, 2.0 Hz), 7.31 (dd,1H, J=11.8, 2.0 Hz), 7.45 (d, 1H, J=8.3 Hz), 7.85 (dd, 1H, J=8.0, 3.3Hz), 8.15 (t, 1H, J=9.2 Hz), 8.18 (s, 1H), 8.38 (d, 1H, J=6.0 Hz), 8.62(d, 1H, J=3.3 Hz), 8.90 (s, 1H), 8.98 (s, 1H), 12.93 (bs, 1H). LC-MS(m/z): 529.12 (M+H, 100). HRMS: m/z calcd. for C27H25FN8O (M+H, 100):529.2106. Found: 529.2095.

(XI) Urea Formation Via Curtius Rearrangement Synthesis 179

Ethyl 3-tert-butyl-1-(cyclopropylmethyl)-1H-pyrazole-5-carboxylate

Method I: A mixture of 3-tert-butyl-1H-pyrazole-5-carboxylate (993 mg,5.06 mmol) caesium carbonate (2.71 g, 8.32 mmol) in dry DMF (10 mL)under Ar was treated dropwise over 15 min with bromomethylcyclopropane(500 μl, 5.16 mmol) at 0° C. The mixture was then allowed to warm up toRT and stirred for 5 h. The mixture was poured into water and extractedwith Et₂O. The combined organic fraction was washed with H₂O, dried(MgSO₄), filtered and evaporated to give residue, which was subsequentlyColumn eluent: 40→100% CH₂Cl₂ in hexane. Yield: 1.10 g (87%) of acolorless oil. Five hours reaction time. ¹H-NMR (DMSO-d₆), δ (ppm), J(Hz): 0.32 (m, 2H, Hcyclopropyl), 0.44 (m, 2H, Hcyclopropyl), 1.25 (m,10H, tert-Bu+Hcyclopropyl), 1.29 (t, J=7.1, 3H, CH₃), 4.28 (m, 4H,NCH₂+OCH₂), 6.71 (s, 1H, H_(arom)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz):3.2, 11.7, 14.0, 30.2, 31.6, 54.8, 60.6, 107.1, 131.4, 159.3, 159.4;LC-MS (m/z): 251.1 (M+H, 100), rt=2.92 min.

Synthesis 180 Ethyl3-tert-butyl-1-(methoxymethyl)-1H-pyrazole-5-carboxylate

Method I was used with ethyl 3-tert-butyl-1H-pyrazole-5-carboxylate(1078 mg, 5.49 mmol), caesium carbonate (2.89 g, 8.87 mmol) andchloro(methoxy)methane (426 μl, 5.60 mmol). 16 hours reaction time.Column eluent: 0→10% EtOAc in CH₂Cl₂. Yield: 485 mg (37%) of a colorlessoil.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.26 (s, 9H, tert-Bu), 1.30 (t, 3H,J=7.1, CH₃), 3.22 (s, 3H, OCH₃), 4.29 (q, 2H, J=7.1, OCH₂CH₃), 5.64 (s,2H, OCH₂N), 6.69 (s, 1H, H_(arom)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz):14.0, 30.0, 31.7, 56.0, 60.8, 80.0, 108.7, 132.7, 158.9, 160.5; LC-MS(m/z): 241.1 (M+H, 100), rt=2.67 min.

Synthesis 181 Ethyl3-tert-butyl-1-(cyclobutylmethyl)-1H-pyrazole-5-carboxylate

Method I: was used with ethyl 3-tert-butyl-1H-pyrazole-5-carboxylate(1085 mg, 5.53 mmol), caesium carbonate (2.89 g, 8.87 mmol) and(bromomethyl)cyclobutane (634 μl, 5.64 mmol). 16 hours reaction time.Column eluent: 40→100% CH₂Cl₂ in hexane. Yield: 0.98 g (67%) of acolorless oil.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.23 (s, 9H, tert-Bu), 1.29 (t, 3H,J=7.1, CH₃), 1.76 (m, 4H, CH₂), 1.89 (m, 2H, CH₂), 2.69 (sept, 1H,J=7.1, CH), 4.27 (q, 2H, J=7.1, OCH₂CH₃), 4.45 (d, 2H, J=7.1, NCH₂),6.69 (s, 1H, H_(arom)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 14.0, 17.7,24.9, 30.2, 31.6, 35.7, 54.9, 60.5, 106.9, 131.6, 159.3 (two coincidentpeaks); LC-MS (m/z): 265.1 (M+H, 100), rt=3.06 min.

Synthesis 182 Ethyl3-tert-butyl-1-(2-(dimethylamino)ethyl)-1H-pyrazole-5-carboxylate

Method I was used with ethyl 3-tert-butyl-1H-pyrazole-5-carboxylate (124mg, 0.632 mmol), caesium carbonate (624 mg, 1.915 mmol) and2-chloro-N,N-dimethylethanamine hydrochloride (96.8 mg, 0.672 mmol). 48hours reaction time. Column eluent: 50→100% EtOAc in CH₂Cl₂, followed by0→10% MeOH in EtOAc. Yield: 103 mg (61%) of a colorless oil.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.25 (s, 9H, tert-Bu), 1.31 (t, 3H,J=7.1, CH₃), 2.15 (s, 6H, N(CH₃)₂), 2.60 (t, 2H, J=6.9, CH₂CH₂NMe₂),4.29 (q, 2H, J=7.1, OCH₂CH₃), 4.51 (t, 2H, J=6.9, CH₂CH₂NMe₂), 6.70 (s,1H, H_(arom)); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 14.1, 30.2, 31.6,45.1, 48.8, 58.7, 60.6, 107.0, 132.1, 159.2, 159.6; LC-MS (m/z): 268.2(M+H, 100), 1.89 min.

Synthesis 1833-Tert-butyl-1-(cyclopropylmethyl)-1H-pyrazole-5-carboxylic acid

Method J: Ethyl3-tert-butyl-1-(cyclopropylmethyl)-1H-pyrazole-5-carboxylate (1.1 g,4.39 mmol) was dissolved in a 4:1:1 mixture of THF/MeOH/H₂O (total 25 mLM), lithium hydroxide monohydrate (200 mg, 4.7 mmol) was added and thecolorless mixture was stirred for 16 h at RT. The volatiles weresubsequently evaporated, the resulting solid was redissolved in H₂O andthe pH of the solution was adjusted to 1 with 10% aqueous HCl. Theresulting milky mixture was extracted with EtOAc and the combinedorganic fraction was washed with brine, dried and concentrated todryness to give a white crystalline solid. Yield: 0.82 g (84%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 0.32 (m, 2H, Hcyclopropyl), 0.42 (m,2H, Hcyclopropyl), 1.24 (m, 10H, tert-Bu+Hcyclopropyl), 4.29 (d, 2H,J=7.0, NCH₂), 6.66 (s, 1H, H_(arom)), 13.10 (br s, 1H, COOH); ¹³C-NMR(DMSO-d₆), δ (ppm), J (Hz): 3.2, 11.8, 30.2, 31.6, 54.6, 107.1, 132.3,159.2, 160.8; LC-MS (m/z): 223.1 (M+H, 100), rt=2.57 min.

Synthesis 184 3-Tert-butyl-1-(methoxymethyl)-1H-pyrazole-5-carboxylicacid

Method J was used with ethyl3-tert-butyl-1-(methoxymethyl)-1H-pyrazole-5-carboxylate (485 mg, 2.02mmol) as the starting material. Yield: 413 mg (96%) of a white,crystalline solid.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.25 (s, 9H, tert-Bu), 3.21 (s, 3H,OCH₃), 5.64 (s, 2H, OCH₂N), 6.79 (s, 1H, H_(arom)), 13.30 (br s, 1H,COOH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 30.1, 31.6, 56.0, 79.7,108.7, 133.8, 160.3, 160.4; LC-MS (m/z): 213.1 (M+H, 100), rt=2.31 min.

Synthesis 185 3-Tert-butyl-1-(cyclobutylmethyl)-1H-pyrazole-5-carboxylicacid

Method J was used with ethyl3-tert-butyl-1-(cyclobutylmethyl)-1H-pyrazole-5-carboxylate (0.98 g,3.71 mmol) as the starting material. Yield: 842 mg (95%) of a white,crystalline solid.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.23 (s, 9H, tert-Bu), 1.76 (m, 4H,CH₂), 1.89 (m, 2H, CH₂), 2.69 (sept, 1H, J=7.1, CH), 4.27 (q, 2H, J=7.1,OCH₂CH₃), 4.45 (d, 2H, J=7.1, NCH₂), 6.64 (s, 1H, H_(arom)), 13.07 (brs, 1H, COOH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 17.7, 24.9, 30.2,31.5, 35.8, 54.7, 106.9, 132.6, 159.1, 160.8 LC-MS (m/z): 237.1 (M+H,100), rt=2.74 min.

Synthesis 1862-(3-Tert-butyl-5-carboxy-1H-pyrazol-1-yl)-N,N-dimethylethanaminiumchloride

Ethyl 3-tert-butyl-1-(2-(dimethylamino)ethyl)-1H-pyrazole-5-carboxylate(98 mg, 0.367 mmol) was dissolved in 6M aqueous HCl (4 mL, 24.00 mmol)and the colorless solution was heated to 80° C. for 72 h. The volatileswere evaporated in vacuo and the resulting white solid was coevaporatedwith Et₂O 10 mL) to give the title compound as a white solid. Yield: 100mg (99%).

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.26 (s, 9H, tert-Bu), 2.78 (d, 6H,J=4.8, CH₂CH₂N+H(CH₃)₂), 3.50 (q, 2H, J=5.3, CH₂CH₂NMe₂), 4.82 (t, 2H,J=6.6, CH₂CH₂NMe₂), 6.78 (s, 1H, H_(arom)), 10.66 (br s, 1H, COOH);LC-MS (m/z): 240.2 (M+H, 100), rt=1.56 min.

Synthesis 1871-(3-Tert-butyl-1-(cyclopropylmethyl)-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-097)

Method F5: 3-tert-butyl-1-(cyclopropylmethyl)-1H-pyrazole-5-carboxylicacid (51 mg, 0.229 mmol) was put under Ar and dry triethylamine (30 uL,0.23 mmol) and dry DMF (1 mL) were subsequently added. The mixture wascooled to 0° C., DPPA (1 equiv) was added at once and the solution wasstirred at 0° C. for an additional 30 min and then at RT for 1 h. Then,8-(4-amino-3-fluorophenoxy)pyrido[2,3-b]pyrazin-3(4H)-one (31.9 mg,0.117 mmol) was added at once and the solution was heated to 100° C. for45 min. The resulting yellow solution was subsequently cooled to RT,diluted with EtOAc. The organic layer was washed with H₂O, 0.1 M citricacid, saturated aqueous NaHCO₃, brine, dried and concentrated to drynessto give a yellow solid. Et₂O was added and the mixture was sonicated for10 min and left to stand. The precipitate was filtered off and washedwith Et₂O to give the desired urea. Yield: 35 mg (62%) of a white solid.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 0.35 (m, 2H, Hcyclopropyl), 0.47 (m,2H, Hcyclopropyl), 1.24 (m, 10H, tert-Bu+Hcyclopropyl), 3.84 (d, 2H,J=6.7, NCH₂), 6.12 (s, 1H, H_(Pyrazole)), 6.66 (d, J=5.6, 1H, H_(Py)),7.07 (m, 1H, H_(arom)), 7.34 (m, 1H, H_(arom)), 8.20 (m, 2H, H_(arom)),8.38 (d, J=5.6, 1H, H_(Py)), 8.80 (br s, 1H, NH), 8.85 (br s, 1H, NH),12.93 (br s, 1H, NH); ¹⁹F-NMR (DMSO-d₆), δ (ppm): −125.0; LC-MS (m/z):492.1 (M+H, 100), 2.54 min; HRMS (3.10 min): m/z calcd. for C₂₅H₂₇FN₇O₃(M+H, 100)⁺: 492.21539. Found: 492.21664.

Synthesis 1881-(3-Tert-butyl-1-(methoxymethyl)-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-098)

Method F5 was employed, using3-tert-butyl-1-(methoxymethyl)-1H-pyrazole-5-carboxylic acid (49.5 mg,0.233 mmol) and8-(4-amino-3-fluorophenoxy)-pyrido[2,3-b]pyrazin-3(4H)-one (31.9 mg,0.117 mmol). Yield: 45 mg (80%) of a white solid.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.24 (s, 9H, tert-Bu), 3.25 (s, 3H,OCH₃), 5.28 (s, 2H, OCH₂N), 6.26 (s, 1H, H_(arom)), 6.67 (d, J=5.6, 1H,H_(Py)), 7.07 (m, 1H, H_(arom)), 7.34 (m, 1H, H_(arom)), 8.22 (m, 2H,H_(arom)), 8.38 (d, J=5.6, 1H, H_(Py)), 9.01 (br s, 1H, NH), 9.11 (br s,1H, NH), 12.93 (br s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 30.1,31.8, 55.7, 77.5, 93.1, 106.5, 108.5 (d, J_(FC)=22.9), 116.5 (d,J_(FC)=3.3), 118.4, 121.5, 124.9 (d, J_(FC)=12.0), 137.6, 145.5, 148.5(d, J_(FC)=10.4), 150.9, 151.2, 152.2, 152.3 (d, J_(FC)=245), 156.5,159.9, 160.5; LC-MS (m/z): 482.1 (M+H, 100), 2.48 min; HRMS (3.05 min):m/z calcd. for C₂₃H₂₄FN₇NaO₄ [M+Na]⁺: 504.17660. Found: 504.17641.

Synthesis 1891-(3-Tert-butyl-1-(cyclobutylmethyl)-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-099)

Method F5 was employed, using3-tert-butyl-1-(cyclobutylmethyl)-1H-pyrazole-5-carboxylic acid (78.5mg, 0.332 mmol) and8-(4-amino-3-fluorophenoxy)-pyrido[2,3-b]pyrazin-3(4H)-one (41 mg, 0.151mmol). Yield: 50 mg (60%) of a white solid.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.22 (s, 9H, tert-Bu), 1.82 (m, 4H,CH₂), 1.98 (m, 2H, CH₂), 2.72 (sept, 1H, J=7.1, CH), 3.96 (d, 2H, J=7.1,NCH₂), 6.11 (s, 1H, H_(arom)), 6.66 (d, J=5.6, 1H, H_(Py)), 7.07 (m, 1H,H_(arom)), 7.33 (m, 1H, H_(arom)), 8.21 (m, 2H, H_(arom)), 8.38 (d,J=5.6, 1H, H_(Py)), 8.79 (br s, 1H, NH), 8.83 (br s, 1H, NH), 12.93 (brs, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 16.4, 23.9, 29.0, 30.4,33.7, 50.5, 91.7, 105.2, 107.1 (d, J_(FC)=22.4), 115.2, 117.0, 120.2,123.7 (d, J_(FC)=10.7), 134.8, 144.2, 147.1 (d, J_(FC)=10.3), 149.8,150.0, 150.8, 150.9 (d, J_(FC)=245), 155.1, 157.3, 159.2 LC-MS (m/z):507.1 (M+H, 100), 2.65 min; HRMS (3.24 min): m/z calcd. forC₂₆H₂₈FN₇NaO₃ [M+Na]⁺: 528.21299. Found: 528.21311.

Synthesis 1901-(3-Tert-butyl-1-(2-(dimethylamino)ethyl)-1H-pyrazol-5-yl)-3-(2-fluoro-4-(3-oxo-3,4-dihydropyrido[3,2-b]pyrazin-8-yloxy)phenyl)urea(AA-100)

Method F5 was employed, using3-tert-butyl-1-(2-(dimethylamino)ethyl)-1H-pyrazole-5-carboxylic acidhydrochloride (89 mg, 0.323 mmol) and8-(4-amino-3-fluorophenoxy)-pyrido[2,3-b]pyrazin-3(4H)-one (41 mg, 0.151mmol). Two equiv of triethylamine were used and the citric acid wash wasnot performed. Yield: 34 mg (41%) of an orange solid.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): 1.21 (s, 9H, tert-Bu), 2.24 (s, 6H,CH₂CH₂N(CH₃)₂), 2.68 (t, 2H, J=6.8, CH₂CH₂NMe₂), 4.04 (t, 2H, J=6.8,CH₂CH₂NMe₂), 6.10 (s, 1H, _(Py)zH), 6.64 (d, J=5.6, 1H, _(Py)rH), 7.06(m, 1H, H_(arom)), 7.33 (m, 1H, H_(arom)), 8.16 (m, 2H, H_(arom)), 8.37(d, J=5.6, 1H, _(Py)rH), 8.89 (br s, 1H, NH), 9.05 (br s, 1H, NH), 12.92(br s, 1H, NH); ¹³C-NMR (DMSO-d₆), δ (ppm), J (Hz): 30.3, 31.8, 45.0,45.5, 57.8, 93.7, 106.5, 108.5 (d, J_(FC)=22.4), 116.4, 118.4, 122.1,124.9 (d, J_(FC)=10.7), 136.6, 145.5, 148.7 (d, J_(FC)=10.3), 151.2,151.6, 152.2, 152.5 (d, J_(FC)=245), 156.5, 159.0, 160.5; ¹⁹F-NMR(DMSO-d₆), δ (ppm): −124.5; LC-MS (1.90 min): m/z 509.1 (M+H, 100); HRMS(3.24 min): m/z calcd. for C₂₅H₃₀FN₈O₃ (M+H, 100)⁺: 509.24194. Found:509.24249.

Synthesis 1915-[(4-amino-2-fluorophenyl-oxy)carbonylamino-5-(1-N-allyl-3-t-butyl-imidazolyl)]-pyridin-[2,3]-3-pyrazin-2-one (AA-095)

Method F2 was used with 34 mg (0.13 mmol) of5-(4-amino-2-fluoro-phenyl-oxy)-pyridin-[2,3]-pyrazin-2-one, and 0.26mmol of 1-N-allyl-3-t-butyl-imidazolyl-5-isocyanate, 38 mg (yield, 42%)of the desired product were obtained.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): δ 4.59 (d, 2H, J=2.5 Hz), 4.90 (d,1H, J=18.6 Hz), 5.15 (d, 1H, J=10.3 Hz), 5.92-6.00 (m, 1H), 6.15 (s,1H_(pyz)), 6.64 (d, 1H, J=5.7 Hz), 7.05 (d, 1H, J=8.6 Hz), 7.34 (d, 2H,J=11.7 Hz), 8.18 (s, 1H_(pyrazine)), 8.20 (t, 1H, J=9.1 Hz), 8.37 (d,1H, J=5.7 Hz), 8.81 (s, 1H, NH), 8.86 (s, 1H, NH), 12.95 (s, 1H, NH).LC-MS (m/z): m/z: 478.1 (M+H, 100)⁺, rt=2.51 min; HRMS: (M+Na)⁺ calcdfor C₂₄H₂₄FN₇O₃Na, 500.1817. Found: 500.1816.

Synthesis 1925-[(4-amino-2-fluorophenyl-oxy)carbonylamino-5-(1-N-propargyl-3-t-butyl-imidazolyl)]-pyridin-[2,3]-3-pyrazin-2-one(AA-096)

Method F2 was used with 35 mg (0.13 mmol) of5-(4-amino-2-fluoro-phenyl-oxy)-pyridin-[2,3]-pyrazin-2-one, and 0.2mmol of 1-N-propargyl-3-t-butyl-imidazolyl-5-isocyanate, 49 mg (yield,80%) of the desired product were obtained.

¹H-NMR (DMSO-d₆), δ (ppm), J (Hz): δ 4.82 (s, 2H), 6.15 (s,1H_(pyrazole)), 6.65 (d, 1H, J=5.7 Hz), 7.06 (d, 1H, J=8.7 Hz), 7.35 (d,2H, J=11.7 Hz), 8.18 (s, 1H_(pyrazine)), 8.20 (t, 1H, J=9.1 Hz), 8.37(d, 1H, J=5.7 Hz), 8.91 (s, 1H, NH), 9.02 (s, 1H, NH), 12.95 (s, 1H,NH). LC-MS (m/z): 476.1 (M+H, 100), rt=2.41 min; HRMS: (M+H, 100)+ calcdfor C₂₄H₂₂FN₇O₃ 476.1841. Found: 476.1844.

Biological Methods Biological Methods—DELFIA Kinase Assay

Compounds were assessed by a kinase assay performed according to thefollowing protocol.

The following reagents were prepared:

DELFIA Kinase Buffer (DKB):

Volume per Volume per Stock mL 10 mL plate Reagent Concentration (μL)(μL) 20 mM MOPS pH 7.2 0.2M 100 1000 0.5M EGTA pH 8.0 0.5M 10 100 10 mMMgCl₂  1M 10 100 0.1% β-mercaptoethanol — 1 10 25 mM β-glycerophosphate0.5M 50 500 Water 100% 829 8290 MOPS = 3-[N-Morpholino] propanesulfonicacid (Sigma M3183). EGTA = Ethyleneglycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (Sigma E3889).DKB1 (DKB with B-RAF and MEK Protein):

Combine 4950 μL of DKB and 50 μL of 2.5 mg/ml GST-MEK stock (to give 1mg of MEK per 40 μL). Then add 22.5 μL of B-RAF to give ˜0.2 μL of B-RAFper 40 μL.

DKB2 (DKB with MEK Protein):

Combine 4950 μL of DKB and 50 μL of 2.5 mg/ml GST-MEK stock (to give 1mg of MEK per 40 μL). Use 500 μL of this for the blow out (BO) and theempty vector (EV) control.

ATP:

100 mM stock, dilute to 500 μM to give 100 μM final concentration inassay.

Inhibitors (Test Compounds):

100 mM stock, dilute to 10, 3, 1, 0.3, 0.1, 0.03, 0.01, 0.003, 0.001,0.0003, 0.0001 mM in DMSO in drug plate, resulting in concentration of100, 30, 10, 3, 1, 0.3, 0.1, 0.03, 0.01, 0.003, 0.001 μM in the assay.

Primary Antibody:

Phospho-MEK1/2 CST #9121S diluted 1:1000 in DELFIA assay buffer (AB).Preincubate antibody in the AB for 30 minutes at room temperature priorto use.

Secondary Antibody:

Anti-rabbit-Eur labelled secondary Perkin Elmer #AD0105 diluted 1:1000in DELFIA assay buffer (AB). Preincubate antibody in the AB for 30minutes at room temperature prior to use. (Primary and secondaryantibodies were incubated together.)

Tween:

0.1% Tween 20 in water.

Assay Buffer:

DELFIA assay buffer Perkin Elmer #4002-0010.

Enhancement Solution:

DELFIA enhancement solution Perkin Elmer #4001-0010.

Assay Plates:

96 well glutathione-coated black plate Perbio #15340.

Procedure:

1. Preblock wells with 5% milk in TBS for 1 hour.2. Wash wells with 3× with 200 μL TBS.3. Plate out 40 μL of DKB1 for all inhibitors (test compounds), DMSOcontrol, and optionally other control compounds.4. Plate out 40 μL of DKB2 for BO and EV wells.5. Add inhibitors (test compounds) at 0.5 μL per well according todesired plate layout.6. Add 0.5 μL DMSO to vehicle control wells.7. Add 2 μL of B-RAF to BO and EV wells.8. Pre-incubate with inhibitors (test compounds) for 10 minutes at roomtemperature with shaking.9. Add 10 μL of 500 μM ATP stock, in DKB, to give 100 μM assayconcentration.10. Seal plates with TopSeal and incubate at room temperature withshaking for 45 minutes.11. Wash plates 3× with 200 μL 0.1% Tween20/Water to terminate reaction.12. Add 50 μL per well of antibody mix and incubate for 1 hour at roomtemperature with shaking.13. Wash plates 3× with 200 μL 0.1% Tween20/Water.14. Add 100 μL DELFIA enhancement solution per well, cover in foil, andincubate at room temperature for 30 minutes with shaking.15. Read on Victor using Europium protocol.

Values for the blank (Empty Vector) are subtracted from all values. TheDMSO controls are set as 100% activity and assay points (the response)are calculated as a percentage of the DMSO control. Data are plottedusing Graphpad Prism software and a non-linear regression line iscalculated using a variable slope sigmoidal dose-response equation(Y=Bottom+(Top−Bottom)/(1+10̂((LogEC50−X)*HillSlope)) where X is thelogarithm of concentration. Y is the response). The IC50 generated bythis procedure is the concentration of the drug that produces apercentage control fluorescence value midway between the saturation, andzero-effect plateaus. Three independent assays are usually performed andthe mean IC50 is reported.

Biological Methods—Cell Based Phosho-ERK Assay

Compounds were assessed using a cell-based assay which was performedaccording to the following protocol.

Day 0:

Plate out 16,000 cells/well in 99 μL medium in a 96-well plate.

Day 1:

1. Add 1 μL inhibitor to the cells (total 1 μL solution).2. Incubate the cells with test compound for 6 hours at 37° C.3. Aspirate off the solution from all of the wells.4. Fixate the cells with 100 μL 4% formaldehyde/0.25% Triton X-100 PBSper well.5. Incubate the plate for 1 hour at 4° C.6. Aspirate off the fixing solution and add 300 μL TBS per well.7. Leave the plate overnight at 4° C.

Day 2:

1. Wash the plate 2× with 200 μL PBS per well.2. Block with 100 μL 5% dried milk in TBS.3. Incubate the plate for 20 minutes at 37° C.4. Wash the plate 2× with 0.1% tween/H₂O.5. Add 50 μL of 3 μg/mL primary antibody pERK (Sigma M8159), diluted in5% milk powder/TBS, to each well.6. Incubate the plate for 2 hours at 37° C.7. Wash the plate 3× with 0.1% tween/H₂O.8. Add 50 μL of 0.45 μg/mL secondary Europium-labelled anti-mouseantibody (Perkin Elmer) to each well.9. Incubate the plate for 1 hour at 37° C.10. Wash the plate 3× with 0.1% tween/H₂O.11. Add 100 μL enhancement solution (Perkin Elmer) to each well.12. Leave the plate for approximately 10 minutes at room temperaturebefore gently shaking the plate.

13. Read Europium Time Resolved Fluorescence in Victor2.

14. Wash the plate 2× with 0.1% tween/H₂O.15. Measure the protein concentration with BCA (Sigma) by adding 200 μLof solution per well.16. Incubate the plate for 30 minutes at 37° C.17. Read absorbance levels at 570 nm in a plate reader.

Note that Europium counts are normalised for protein levels by dividingcounts by absorbance.

Values for the blank (no cells) are subtracted from all values. The DMSOcontrols are set as 100% activity and assay points (the response) arecalculated as a percentage of the DMSO control. Data are plotted usingGraphpad Prism software and a non-linear regression line is calculatedusing a variable slope sigmoidal dose-response equation(Y=Bottom+(Top−Bottom)/(1+10̂((LogEC50−X)*HillSlope)) where X is thelogarithm of concentration. Y is the response). The IC50 generated bythis procedure is the concentration of the drug that produces apercentage control fluorescence value midway between the saturation, andzero-effect plateaus. Three independent assays are usually performed andthe mean IC50 is reported.

Biological Methods —SRB Cell Proliferation Assay (SRB GI₅₀)

Cultures of WM266.4 melanoma cells are routinely cultured in DMEM/10%foetal bovine serum, at 37° C., in 5% CO₂ water saturated atmosphere.Cultures are maintained in exponential growth phase by sub-culturingbefore having become confluent (3-5 day intervals). Single cellsuspensions are prepared by harvesting an 80 cm² tissue culture flaskwith 5 mL commercial trypsin EDTA. After 5 minutes, the detached cellsare mixed with 5 mL fully complemented culture medium and centrifugallypelleted (1000 rpm for 7 minutes). After aspirating the supernatant, thecell pellet is re-suspended in 10 mL fresh medium and the cells fullydisaggregated by drawing the whole volume up/down 5 times through a19-gauge needle. The concentration of the cells is determined using ahaemocytometer (1/10 dilution). A suitable volume to give at least a2-fold excess for the number of tests being conducted, typically 100-200mL, is prepared by diluting the cell suspension to 10,000/mL, and 100μL/well dispensed into 96 well plates using a programmable 8-channelperistaltic pump, giving 1000 cells/well, leaving column 12 blank. Theplates are returned to the incubator for 24 hours to allow the cells tore-attach.

The compounds being tested are prepared at 20 mM in dimethylsulphoxide.Aliquots (200 μL) are diluted into 20 mL culture medium giving 200 μM,and 10 serial dilutions of 3× performed by transferring 5 mL to 10 mL.Aliquots (100 μL) of each dilution are added to the wells, using an8-channel pipettor, thus performing a final further 2× dilution, andgiving doses ranging from 100 μM to 0.005 μM. Column 11 receives plainculture medium only. Each compound is tested in quadruplicate, eachreplicate being the average of four wells, and two plates per compound.

After a further 6 days growth, the plates are emptied, and the cells arefixed in 10% trichloroacteic acid for 10 minutes on ice. After thoroughrinsing in running tap water, the plates are dried, and stained byadding 50 μL of a solution of 0.1% sulphorhodamine-B in 1% acetic acid,for 10 minutes at room temperature. The stain is poured out and theplates thoroughly rinsed under a stream of 1% acetic acid, thus removingunbound stain, and dried. The bound stain is taken into solution byaddition of 150 μL Tris buffer pH 8, followed by 10 minutes on aplate-shaker (approximately 500 rpm). The absorbance at 540 nm in eachwell (being proportional to the number of cells present) is determinedusing a plate reader.

After averaging the results in rows A-D and E-H, the blank value (row12) is subtracted, and results expressed as percentage of the untreatedvalue (row 11). The 10 values so derived (in quadruplicate) are plottedagainst the logarithm of the drug concentration, and analysed bynon-linear regression to a four parameter logistic equation, settingconstraints if suggested by inspection. The GI₅₀ generated by thisprocedure is the concentration of the drug that produces a percentagecontrol A₅₄₀ midway between the saturation, and zero-effect plateaus.

Biological Results

The following compounds were tested in the “DELFIA Kinase Assay”described above: AA-001 through AA-056.

The following compounds have an IC50 BRAF of less than 1.0 μM:

AA-001, AA-002, AA-003, AA-004, AA-005, AA-006, AA-007, AA-008, AA-009,AA-010, AA-011, AA-012, AA-013, AA-014, AA-015, AA-016, AA-017, AA-018,AA-019, AA-020, AA-021, AA-022, AA-023, AA-024, AA-025, AA-026, AA-027,AA-028, AA-029, AA-030, AA-031, AA-032, AA-034, AA-037, AA-038, AA-039,AA-042, AA-044, AA-045, AA-046, AA-047, AA-048, AA-049, AA-050, AA-051,AA-052, AA-053, AA-054, AA-055, AA-056.

Additionally, the following compounds were tested in the “DELFIA KinaseAssay” described above: AA-001 through AA-098.

The following compounds have an IC50 BRAF of less than 0.1 μM:

AA-002, AA-003, AA-004, AA-005, AA-006, AA-007, AA-008, AA-009, AA-010,AA-011, AA-014, AA-015, AA-017, AA-018, AA-019, AA-020, AA-021, AA-023,AA-024, AA-025, AA-026, AA-027, AA-028, AA-029, AA-032, AA-044, AA-045,AA-047, AA-048, AA-050, AA-051, AA-052, AA-054, AA-060, AA-061, AA-062,AA-063, AA-064, AA-065, AA-067, AA-069, AA-072, AA-074, AA-075, AA-079,AA-080, AA-086, AA-087, AA-088, AA-093, AA-094, AA-095, AA-096, AA-097,AA-098.

The following compounds have an IC50 BRAF of at least 0.1 μM and lessthan 1.0 μM:

AA-001, AA-012, AA-013, AA-016, AA-022, AA-030, AA-031, AA-033, AA-034,AA-035, AA-037, AA-038, AA-039, AA-040, AA-041, AA-042, AA-043, AA-046,AA-049, AA-053, AA-055, AA-056, AA-057, AA-058, AA-059, AA-066, AA-068,AA-071, AA-076, AA-077, AA-078, AA-081, AA-082, AA-083, AA-084, AA-085,AA-089, AA-090, AA-091, AA-092.

One compound, compound AA-016, has an IC50 BRAF of 0.252 μM.

The following compounds were tested in the “Cell Based Phospho-ERKAssay” described above: AA-001 through AA-056.

The following compounds have an IC50 pERK of less than 10 μM:

AA-001, AA-002, AA-003, AA-004, AA-005, AA-006, AA-007, AA-008, AA-009,AA-010, AA-011, AA-013, AA-014, AA-015, AA-016, AA-017, AA-018, AA-019,AA-020, AA-021, AA-022, AA-023, AA-024, AA-025, AA-026, AA-027, AA-028,AA-029, AA-031, AA-033, AA-034, AA-035, AA-036, AA-037, AA-038, AA-039,AA-040, AA-041, AA-043, AA-044, AA-045, AA-046, AA-050, AA-051, AA-052,AA-053, AA-054.

Additionally, the following compounds were tested in the “Cell BasedPhospho-ERK Assay” described above: AA-001 through AA-099.

The following compounds have an IC50 pERK of less than 1.0 μM:

AA-003, AA-006, AA-008, AA-009, AA-010, AA-011, AA-014, AA-015, AA-016,AA-017, AA-018, AA-019, AA-023, AA-024, AA-025, AA-026, AA-028, AA-031,AA-033, AA-034, AA-035, AA-036, AA-040, AA-041, AA-051, AA-052, AA-053,AA-057, AA-059, AA-060, AA-061, AA-062, AA-063, AA-064, AA-065, AA-066,AA-067, AA-071, AA-072, AA-073, AA-074, AA-075, AA-077, AA-078, AA-079,AA-080, AA-081, AA-084, AA-085, AA-087, AA-088, AA-089, AA-090, AA-091,AA-093, AA-094, AA-095, AA-096, AA-097, AA-099.

The following compounds have an IC50 pERK of at least 1.0 μM and lessthan 10 μM:

AA-001, AA-002, AA-004, AA-005, AA-007, AA-013, AA-020, AA-021, AA-022,AA-027, AA-029, AA-037, AA-038, AA-039, AA-043, AA-044, AA-045, AA-046,AA-050, AA-054, AA-058, AA-069, AA-070, AA-076, AA-083, AA-086, AA-092,AA-098.

One compound, compound AA-016, has an IC50 ppERK of 0.096 μM.

The following compounds were tested in the “SRB Cell ProliferationAssay” described above: AA-001 through AA-036 and AA-038 through AA-056.

The following compounds have a GI50 SRB of less than 10 μM:

AA-001, AA-002, AA-003, AA-004, AA-005, AA-006, AA-008, AA-009, AA-010,AA-011, AA-013, AA-014, AA-015, AA-016, AA-017, AA-018, AA-019, AA-020,AA-021, AA-022, AA-023, AA-024, AA-025, AA-026, AA-027, AA-028, AA-029,AA-030, AA-031, AA-032, AA-033, AA-034, AA-035, AA-036, AA-037, AA-038,AA-039, AA-040, AA-041, AA-042, AA-043, AA-044, AA-045, AA-046, AA-047,AA-048, AA-049, AA-050, AA-051, AA-052, AA-053, AA-054, AA-056.

Additionally, he following compounds were tested in the “SRB CellProliferation Assay” described above: AA-001 through AA-036 and AA-038through AA-099.

The following compounds have an GI50 SRB of less than 1.0 μM:

AA-005, AA-006, AA-008, AA-009, AA-010, AA-011, AA-014, AA-015, AA-016,AA-017, AA-018, AA-019, AA-023, AA-024, AA-027, AA-028, AA-031, AA-033,AA-034, AA-035, AA-038, AA-040, AA-041, AA-051, AA-052, AA-053, AA-056,AA-057, AA-059, AA-060, AA-061, AA-062, AA-063, AA-064, AA-065, AA-066,AA-067, AA-071, AA-073, AA-074, AA-075, AA-077, AA-078, AA-079, AA-080,AA-081, AA-084, AA-085, AA-087, AA-088, AA-089, AA-090, AA-091.

The following compounds have an GI50 SRB of at least 1.0 μM and lessthan 10 μM:

AA-001, AA-002, AA-003, AA-004, AA-007, AA-012, AA-013, AA-020, AA-021,AA-022, AA-025, AA-026, AA-029, AA-030, AA-032, AA-036, AA-039, AA-042,AA-043, AA-044, AA-045, AA-046, AA-047, AA-048, AA-049, AA-050, AA-054,AA-055, AA-058, AA-068, AA-069, AA-070, AA-072, AA-076, AA-083, AA-086,AA-092, AA-093, AA-094, AA-095, AA-096, AA-097, AA-098, AA-099.

One compound, compound AA-016, has a GI50 SRB of 0.062 μM.

In Vivo Study 1 AA-018 Non-Established 5 mg/kg/Day Intraperitoneally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.The following day, treatment with test compound was begun. A suspensionof test compound in DMSO:saline for injection 1:19 (v:v) was injectedintraperitoneally at 10 mL/kg bodyweight. Treatment was continued dailyfor 24 doses. The results are shown in FIG. 1.

In Vivo Study 2 AA-018 Non-Established 10 mg/kg/Day Intraperitoneally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.The following day, treatment with test compound was begun. A suspensionin DMSO:saline for injection 1:19 (v:v) was injected intraperitoneallyat 10 mL/kg bodyweight. Treatment was continued daily for 18 doses. Theanimals were then observed after the end of treatment. The results areshown in FIG. 2.

In Vivo Study 3 AA-019 Non-Established 5 mg/kg/Day Intraperitoneally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.The following day, treatment with test compound was begun. A suspensionin DMSO:saline for injection 1:19 (v:v) was injected intraperitoneallyat 10 mL/kg bodyweight. Treatment was continued daily for 24 doses. Theresults are shown in FIG. 3.

In Vivo Study 4 AA-019 Non-Established 10 mg/kg/Day Intraperitoneally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.The following day, treatment with test compound was begun. A suspensionin DMSO:saline for injection 1:19 (v:v) was injected intraperitoneallyat 10 mL/kg bodyweight. Treatment was continued daily for 18 doses. Theanimals were then observed after the end of treatment. The results areshown in FIG. 4.

In Vivo Study 5 AA-019 Non-Established 15 mg/kg/Day Orally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.The following day, treatment with test compound was begun. A suspensionin DMSO:water 1:19 (v:v) was administered by gavage at 10 mL/kgbodyweight. Treatment was continued daily for 24 doses. The results areshown in FIG. 5.

In Vivo Study 6 AA-019 Established 10/5 mg/kg/Day Intraperitoneally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.Groups of 8 from the middle range of tumour sizes were assigned totreatments by stratified allocation on tumour volume. Treatment withtest compound at 10 mg/kg was begun on day 12 after giving cells. Asuspension in DMSO:saline for injection 1:19 (v:v) was injectedintraperitoneally at 10 mL/kg bodyweight. After 10 doses, the dosage wasreduced to 5 mg/kg/day. Treatment was daily for a total of 24 doses. Theresults are shown in FIG. 6.

In Vivo Study 7 AA-019 Established 15 mg/kg/Day Orally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.Groups of 8 from the middle range of tumour sizes were assigned totreatments by stratified allocation on tumour volume. Treatment withtest compound was begun on day 12 after giving cells. A suspension inDMSO:water 1:19 (v:v) was administered by gavage at 10 mL/kg bodyweight.Treatment was continued daily for 24 doses. The results are shown inFIG. 7.

In Vivo Study 8 AA-062 Established 50 mg/kg/Day Orally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.Groups of 8 from the middle range of tumour sizes were assigned totreatments by stratified allocation on tumour volume. Treatment withtest compound was begun on day 13 after giving cells. A suspension inDMSO:water 1:19 (v:v) was administered by gavage at 10 mL/kg bodyweight.Treatment was continued daily for 24 doses. The results are shown inFIG. 8.

In Vivo Study 9 AA-067 Established 10 mg/kg/Day Orally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.Groups of 8 from the middle range of tumour sizes were assigned totreatments by stratified allocation on tumour volume. Treatment withtest compound was begun on day 14 after giving cells. A suspension inDMSO:water 1:19 (v:v) was administered by gavage at 10 mL/kg bodyweight.Treatment was continued daily for 24 doses. The results are shown inFIG. 9.

In Vivo Study 10 AA-017 Established 20 mg/kg/Day Orally

10⁷ A375M human melanoma cells were inoculated sub-cutaneously in 0.2 mLsuspension into the right flank of female Crl:CD1-Foxn1nu athymic mice.Groups of 8 from the middle range of tumour sizes were assigned totreatments by stratified allocation on tumour volume. Treatment withtest compound was begun on day 14 after giving cells. A suspension inDMSO:water 1:19 (v:v) was administered by gavage at 10 mL/kg bodyweight.Treatment is continuing daily for 24 doses (data for the first 16 daysare provided). The results are shown in FIG. 10.

The foregoing has described the principles, preferred embodiments, andmodes of operation of the present invention. However, the inventionshould not be construed as limited to the particular embodimentsdiscussed. Instead, the above-described embodiments should be regardedas illustrative rather than restrictive, and it should be appreciatedthat variations may be made in those embodiments by workers skilled inthe art without departing from the scope of the present invention.

1. A compound selected from compounds of the following formula, andpharmaceutically acceptable salts thereof:

wherein: —R^(Q1) is independently —H, —R¹, —R^(1X), —Cl, —OH, —OR¹,—OR^(1X), —SH, —SR¹, —NH₂, —NHR¹, —NR¹ ₂, or —NR^(RA)R^(RB); wherein:each —R¹ is independently saturated aliphatic C₁₋₆alkyl, and isunsubstituted or substituted with one or more groups selected from —OH,—OR¹¹, —NH₂, —NHR¹¹, and —NR¹¹ ₂, wherein each —R¹¹ is independentlysaturated aliphatic C₁₋₃alkyl; each —R^(1X) is independently saturatedaliphatic C₁₋₄alkyl substituted with one or more groups selected from—F, —Cl, —Br, and —I; and NR^(RA)R^(RB) is independently azetidino,pyrrolidino, piperidino, piperazino, morpholino, azepino, or diazepino,and is optionally substituted with one or more groups selected fromsaturated aliphatic C₁₋₄alkyl; —R^(Q2) is independently —H, —R²,—R^(2X), —Cl, —OH, —OR², —OR^(2X), —SH, —SR², —NH₂, —NHR², —NR² ₂, or—NR^(RC)R^(RD); wherein: each —R² is independently saturated aliphaticC₁₋₆alkyl, and is unsubstituted or substituted with one or more groupsselected from —OH, —OR²², —NH₂, —NHR²², and —NR²² ₂, wherein each —R²²is independently saturated aliphatic C₁₋₃alkyl; each —R^(2X) isindependently saturated aliphatic C₁₋₄alkyl substituted with one or moregroups selected from —F, —Cl, —Br, and —I; and —NR^(RC)R^(RD) isindependently azetidino, pyrrolidino, piperidino, piperazino,morpholino, azepino, or diazepino, and is optionally substituted withone or more groups selected from saturated aliphatic C₁₋₄alkyl; —X— isindependently —O—, —S—, —S(═O)—, or —S(═O)₂—; -M- is independentlyselected from:

wherein: each n is independently 0, 1 or 2; and each R^(PH1) isindependently —F, —Cl, —Br, —I, —R³, —R^(3Y), —CF₃, —OH, —OR³, —OCF₃,—NH₂, —NHR³, —NR³ ₂, —CN, —SH, or —SR³; wherein each —R³ isindependently saturated aliphatic C₁₋₄alkyl, and each —R^(3Y) isindependently aliphatic C₂₋₆alkenyl or aliphatic C₂₋₆alkynyl; J-L- isindependently selected from: J-NR^(N1)—C(═Y)—NR^(N1)—, J-NR^(N1)—C(═Y)—,and J-C(═Y)—NR^(N1)—; wherein: each —R^(N1) is independently —H orsaturated aliphatic C₁₋₄alkyl; and each ═Y is independently ═O or ═S;and -J is independently phenyl or C₅₋₆heteroaryl, and is optionallysubstituted with one or more substituents selected from: —F, —Cl, —Br,—I, —CF₃, —OCF₃, —R⁴, —R^(4S), —R^(4A), —R^(4B), —R^(4C), -L⁴-R^(4C),—Ar, -L⁴-Ar, —OH, —OR¹, -L⁴-OH, -L⁴-OR⁴, —O-L⁴-OH, —O-L⁴-OR⁴, —OR^(4C),—O-L⁴-R^(4C), —OAr, —O-L⁴-Ar, —SH, —SR⁴, —CN, —NO₂, —NH₂, —NHR^(4SS),—R^(N), -L⁴-NH₂, -L⁴-NHR^(4SS), -L⁴-R^(N), —O-L⁴-NH₂, —O-L⁴-NHR^(4SS),—O-L⁴-R^(N), —NH-L⁴-NH₂, —NH-L⁴-NHR^(4SS), —NH-L⁴-R^(N), —NR⁴-L⁴-NH₂,—NR⁴-L⁴-NHR^(4SS), —NR⁴-L⁴-R^(N), wherein: each —R⁴ is independentlysaturated aliphatic C₁₋₆alkyl; each —R^(4S) is independently saturatedaliphatic C₁₋₆alkyl substituted with one or more groups selected from—OH, —OR^(4SS), —C(═O)OH, —C(═O)OR^(4SS), —NH₂, —NHR^(4SS),—N(R^(4SS))₂, —R^(N), —C(═O)NH₂, —C(═O)NHR^(4SS), —C(═O)N(R^(4SS))₂, and—C(═O)R^(N); each —R^(4A) is independently aliphatic C₂₋₆alkenyl; each—R^(4S) is independently aliphatic C₂₋₆alkynyl; each —R^(4C) isindependently saturated C₃₋₆cycloalkyl optionally substituted with oneor more substituents selected from —F, —R⁵, —OH, —OR⁵, —CF₃, and —OCF₃,each -L⁴- is independently saturated aliphatic C₁₋₄alkylene; each —Ar isphenyl or C₅₋₆heteroaryl optionally substituted with one or moresubstituents selected from —F, —Cl, —Br, —I, —R⁵, —OH, —OR⁵, —CF₃,—OCF₃, and —S(═O)₂R⁵; each —R^(4SS) is independently saturated aliphaticC₁₋₄alkyl; each —R^(N) is independently azetidino, pyrrolidino,piperidino, piperazino, morpholino, azepino, or diazepino, and isoptionally substituted with one or more groups selected from saturatedaliphatic C₁₋₄alkyl; and each —R⁵ is independently saturated aliphaticC₁₋₄alkyl.
 2. A compound according to claim 1, wherein: —R^(Q1) isindependently —H, —OH, -Me, —CF₃, —CH₂Br, —NH₂, —NHMe, —NMe₂,morpholino, or piperazino, or N-methyl-piperazino; and —R^(Q2) isindependently —H, —OH, -Me, —CF₃, —CH₂Br, —NH₂, —NHMe, —NMe₂,morpholino, or piperazino, or N-methyl-piperazino.
 3. A compoundaccording to claim 1, wherein: either: —R^(Q1) is —OH, and —R^(Q2) isindependently —H, -Me, —NH₂, —NHMe, morpholino, or piperazino, orN-methyl-piperazino. or: —R^(Q1) is independently —H, -Me, —NH₂, —NHMe,morpholino, or piperazino, or N-methyl-piperazino, and —R^(Q2) is —OH.4. A compound according to claim 1, wherein: either: —R^(Q1) is -Me or—NH², and —R^(Q2) is —OH; or: —R^(Q1) is —OH, and —R^(Q2) is -Me or—NH².
 5. A compound according to claim 1, wherein: either: —R^(Q1) is—OH, and —R^(Q2) is —H; or: —R^(Q1) is —H, and —R^(Q2) is —OH.
 6. Acompound according to claim 1, wherein —X— is independently —O—.
 7. Acompound according to claim 1, wherein -M- is independently:


8. A compound according to claim 1, wherein -M- is independently:

wherein each —R^(PH1) is independently —F, —Cl, —Br, —I, —R³, —OH, —OR³,—SH, or —SR³; wherein each —R³ is independently saturated aliphaticC₁₋₄alkyl.
 9. A compound according to claim 1, wherein J-L- isindependently J-NH—C(═O)—NH—, J-NH—C(═O)—, or J-C(═O)—NH—.
 10. Acompound according to claim 1, wherein J-L- is independentlyJ-NH—C(═O)—NH—.
 11. A compound according to claim 1, wherein -J isindependently phenyl, pyrazolyl, or pyridyl, and is optionallysubstituted with one or more substituents selected from —F, —Cl, —Br,—I, —R⁴, —OH, —OR⁴, —CF₃, —OCF₃, and -Ph; wherein each —R⁴ isindependently saturated aliphatic C₁₋₄alkyl; and each -Ph denotes phenyloptionally substituted with one or more substituents selected from —F,—Cl, —Br, —I, —R⁵, —OH, —OR⁵, —CF₃, and —OCF₃, wherein each —R⁵ isindependently saturated aliphatic C₁₋₄alkyl.
 12. A compound according toclaim 1, wherein -J is independently:

wherein: —R^(PY1) is independently phenyl or pyridyl, and is optionallysubstituted with one or more substituents selected from —F, —Cl, —Br,—I, —R⁵, —OH, —OR⁵, —CF₃, —OCF₃, wherein each —R⁵ is independentlysaturated aliphatic C₁₋₄alkyl; —R^(PY2) is independently —F, —Cl, —Br,—I, —R⁴, —OH, —OR⁴, —CF₃, —OCF₃, and -Ph, wherein each —R⁴ isindependently saturated aliphatic C₁₋₄alkyl.
 13. A compound according toclaim 12, wherein: —R^(PY1) is independently phenyl, and is optionallysubstituted with one or more substituents selected from —F, —Cl, —Br,—I, -Me, —OH, and —OMe; and —R^(PY2) is independently -tBu.
 14. Acompound according to claim 1, wherein -J is independently:

wherein: m is independently 0, 1, or 2; each —R^(PH2) is independently—F, —Cl, —Br, —I, —R⁴, —OH, —OR⁴, —CF₃, or —OCF₃, wherein each —R⁴ isindependently saturated aliphatic C₁₋₄alkyl.
 15. A pharmaceuticalcomposition comprising a compound according to claim 1, and apharmaceutically acceptable carrier or diluent.
 16. A method ofpreparing a pharmaceutical composition comprising the step of admixing acompound according to claim 1, and a pharmaceutically acceptable carrieror diluent.
 17. A method of treatment of a disease or disorder that isameliorated by the inhibition of RAF function, a proliferative disorder,cancer, melanoma, or colorectal cancer comprising administering to asubject in need of treatment a therapeutically-effective amount of acompound according to claim
 1. 18. A method of inhibiting RAF function,in vitro or in vivo, comprising contacting a RAF with an effectiveamount of a compound according to claim
 1. 19. A method of inhibitingRAF function in a cell, in vitro or in vivo, comprising contacting thecell with an effective amount of a compound according to claim
 1. 20. Amethod of inhibiting cell proliferation, inhibiting cell cycleprogression, promoting apoptosis, or a combination of one or more these,in vitro or in vivo, comprising contacting the cell with an effectiveamount of a compound according to claim 1.